Post on 14-Oct-2020
MODE OF INHERITANCE FOR YIELD AND
YIELD RELATED TRAITS IN CHICKPEA
(Cicer arietnium L.)
By
Khalid Mahmood
M.Sc. (Hons.) Agriculture
A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE
REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
IN
PLANT BREEDING AND GENETICS
Department Of Plant Breeding and Genetics
UNIVERSITY OF AGRICULTURE
FAISALABAD
PAKISTAN
2011
To The Controller of Examinations,
University of Agriculture,
Faisalabad.
“We, the Supervisory Committee, certify that the contents and form of thesis
submitted by Mr. Khalid Mahmood , 84-ag-1187, have been found satisfactory and
recommend that it be processed for evaluation, by the External Examiner(s) for the award of
degree”.
Supervisory Committee
1. Chairman ___________________________ (Dr. Muhammad Saleem) 2. Member ___________________________ (Dr. Muhammad Ahsan) 3. Member ___________________________ (Dr. Ashfaq Ahmad Chattha)
CONTENTS
S.No. Subject Page no.
Acknowledgements Chapter-1
1 INTRODUCTION 1
Chapter-2
REVIEW OF LITERATURE 4
1 Gene action studies 4
2 Combining ability studies 9
3 Molecular Studies 15
4 Wilt inheritance studies 16
Chapter-3
MATERIALS AND METHODS 18
1 Genetic Analysis 21
2 Combining Ability Analysis 26
3 Molecular Study 27
4 Chickpea wilt inheritance 29
Chapter-4
RESULTS AND DISCUSSION 31
Analysis of variance 31
4.1. Diallel analysis 31
Number of days taken to flowering 34
Number of primary branches per plant 38
Number of secondary branches per plant 42
Plant Height (cm) 46
Number of days taken to maturity 51
Total weight of plant (g) 55
Number of pods per plant 59
Number of seeds per pod 64
100-seed weight (g) 68
Grain yield per plant (g) 73
4.2 Combining Ability 77
Number of days taken to flowering 77
Number of primary branches per plant 80
Number of secondary branches per plant 82
Plant Height (cm) 84
Number of days taken to maturity 84
Total weight of plant (g) 86
Number of pods per plant 87
Number of seeds per pod 89
100-seed weight (g) 89
Grain yield per plant (g) 91
Harvest index (%) 93
4.3 Molecular Studies 93
4.4 Inheritance of resistance to chickpea wilt. 104
Chapter-5
SUMMARY 106
LITERATURE CITED 109
LIST OF TABLES
S.No. Title Page no.
1 Characteristics of parental genotypes 18
2 Analysis of variance for yield and its various components. 32
3 Adequacy test of additive dominance model for 6×6 diallel cross of Cicer arietinum L.
33
4 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for number of days taken to flowering in chickpea.
35
5 Estimates of genetic components of variation for number of days taken to flowering in chickpea.
36
6 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for number of primary branches per plant in chickpea.
39
7 Estimates of genetic components of variation for number of primary branches per plant in chickpea.
40
8 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for number of secondary branches per plant in chickpea.
43
9 Estimates of genetic components of variation for number of secondary branches in chickpea.
44
10 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for Plant height (cm) in chickpea.
48
11 Estimates of genetic components of variation for plant height (cm) in chickpea.
49
12 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for number of days taking to maturity in chickpea.
52
13 Estimates of genetic components of variation for number of days taken to maturity in chickpea.
53
14 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for total weight of plant (g) in chickpea.
56
15 Estimates of genetic components of variation for total weight of plant in chickpea.
57
16 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for number of pods per plant in chickpea.
61
17 Estimates of genetic components of variation for number of pods per plant in chickpea.
62
18 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for number of seeds per pod in chickpea.
65
19 Estimates of genetic components of variation for number of seeds per pod in chickpea.
66
20 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for 100-seed weight (g) in chickpea.
69
21 Estimates of genetic components of variation for 100-Seed weight (g) in chickpea.
70
22 Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for grain yield per plant (g) in chickpea.
74
23 Estimates of genetic components of variation for grain yield per plant in chickpea.
75
24 Mean squares and significances from the analysis of variance of combining ability in 6x6 diallel cross of chickpea.
78
25 Estimates of variation components, general (σ2g) & specific combing ability (σ2s), reciprocal effects (σ2r), error (σ2e) and GCA/SGA ratio in 6×6 diallel cross of chickpea
79
26 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number days taken to flowering.
81
27 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of primary branches per plant.
81
28 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of secondary branches per plant.
83
29 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for plant height (cm).
83
30 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number days taken to maturity.
85
31 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for total weight of plant (g).
85
32 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of pods per plant.
88
33 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of seeds per pod.
88
34 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for 100-seed weight (g).
90
35 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for grain yield per plant (g).
90
36 General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for harvest index (%).
92
37 Similarity matrix for Nei and Li's Original Measures of Genetic Identity and Genetic distance of 21 Chickpea genotypes
95
38 Number of resistant and susceptible plants or progenies in F1, F2 and F3 generations of the 6 crosses infested with Fusarium oxysporum L.
105
LIST OF FIGURES
S.No. Title Page no.
1 Wr/Vr graph for days taken to flowering 37
2 Vr/Wr graph for number of primary branches per plant 41
3 Vr/Wr graph for number of secondary branches per plant 45
4 Vr/Wr graph for plant height 50
5 Wr/Vr graph for number of days taken to maturity 54
6 Vr/Wr graph for total weight of plant 58
7 Vr/Wr graph for number of pods per plant 63
8 Vr/Wr graph for number of seeds per pod 67
9 Vr/Wr graph for 100-seed weight 71
10 Vr/Wr graph for grain yield per plant 76
11 Amplification profile of 21 Chickpea genotypes with primer GLD-11
96
12 Amplification profile of 21 Chickpea genotypes with primer GLA-01
97
13 Amplification profile of 21 Chickpea genotypes with primer GLD-01
98
14 Amplification profile of 21 Chickpea genotypes with primer GLB-14 99
15 Amplification profile of 21 Chickpea genotypes with primer GLC-20
100
16 Amplification profile of 21 Chickpea genotypes with primer GLA-09
101
17 Amplification profile of 21 Chickpea genotypes with primer GLA-02
102
18 A dendrogram of 21 chickpea genotypes 103
ACKNOWLEDGEMENTS
All praises to Almighty Allah, the most merciful and the most compassionate, Who
enabled me to accomplish this study successfully. After Almighty Allah, all praises are to the
Holy Prophet Hazrat Muhammad (may peace be upon him), the most perfect and exalted
amongst the ever born on the surface of the earth, who is a beacon of guidance for all the
humanity.
I wish to express sincere appreciation to my supervisor Dr. Muhammad Saleem,
Professor, Department of Plant Breeding and Genetics, University of Agriculture Faisalabad,
for his intent concern and throughout this research manuscript. His gentle and tender
disposition always provided me a source of inspiration during the course of the study.
I like to extend my cordial gratitude to Dr. Muhammad Ahsan, Assistant Professor,
Department of Plant Breeding and Genetics, University of Agriculture Faisalabad, for his
keen interest and valuable assistance as and whenever needed. I am highly obliged to Dr.
Ashfaq Ahmad Chattha, Associate Professor, Department of Agronomy for his cooperation
and valuable suggestions during the course of study.
I pay special gratitude to Dr. Abdus Salam Khan, Professor and Chairman, Department of
Plant Breeding and Genetics, University of Agriculture Faisalabad, for valuable and critical
perusal of the manuscript.
Cordial thanks are also extended to my friends, Ghulam Nabi and Rashid Mahmood for
their excellent cooperation.
I wish to record special thanks to my family father, mother and wife. Their heartfelt
blessings have always been a major source for my accomplishment. Their moral and
financial support and encouragement always helped me for the successful completion of my
studies.
Khalid Mahmood
Abbreviations
Some abbreviations used throughout the text are as follow
AGRIC = Agricultural
ANN = Annals
APPL = Applied
BIO = Biology
d.f = Degree of freedom
F1 = First filial generation
F2 = Second filial generation
F3 = Third filial generation
1
Chapter-1
INTRODUCTION
Chickpea (Cicer arietnium L.), a cool season food legume grown in tropical, sub-
tropical and temperate regions of the world is called by various names i.e., Bengal gram
(India), chana (Urdu), chickpea and gram (English), Garbanzo (Latin America), Hommas,
Hamaz (Arab world), Nohud, Lablabi (Turkey) and Shimbra (Ethiopia). Kabuli types are
predominant in the temperate regions while desi type chickpea is grown in the semi-arid
tropics. Chickpea is valued for its nutritive seeds with high protein contents (20.0-24.0%),
38-59% carbohydrate, 3% fiber, 4.8-5.5% oil and 3% ash. Raw whole seeds contain 357
calories per 100 g, 4.5-15.69% moisture, 14.9-24.6 g protein, 2-4.8 g ash, 140-440 mg Ca,
190-383 mg P, 5.0-23.9 mg Fe, 0.12-0.33 mg riboflavin and 1.3-2.9 mg niacin (Huisman
and Poel, 1994).
The major chickpea growing countries are India, Pakistan and Turkey in Asia,
Ethiopia in Africa, California and Washington states in the USA, Mexico and Australia. In
Pakistan, the crop is grown under three farming systems: the rainfed system, contributing
88% of the total chickpea area, where it is grown as sole crop or mixed with others; the rice
based system constituting 11% of the total chickpea area, where the crop is grown on
residual moisture after rice and irrigated system constituting 1% of the total area. The area
under the crop is about 1028.9 thousand ha as winter crop with a production of 479.5
thousand tones (Anonymous, 2008-09). The major chickpea production belt is the ‘Thal’
comprising the districts of Bhakhar, Mianwali, Leyyah, Khushab and parts of Jhang as
well as Attock, Rawalpindi, Jehlum and Chakwal districts of Potohar. Since most of the
chickpea crop is grown under the rainfed system, fluctuations in rainfall can severely limit
productivity.
The two largest chickpea growing countries i.e., India and Pakistan, which account
for nearly 85% of the chickpea area in the world have low average yield levels of 598 kg/ha
and 466 kg/ha, respectively. The low yield of chickpea in countries with extensive area
under the crop is the factor responsible for overall low production of the crop in the world.
This is primarily due to paucity of productive cultivars, non-availability of good quality
2
seed and resistance against various biotic and abiotic stresses. Of the various fungal
diseases, chickpea wilt caused by Fusarium oxysporum sp. ciceris causing the plant to wilt,
is a serious problem especially in the rainfed area. Therefore, introduction and evolution of
chickpea genotypes performing better in both rainfed and irrigated conditions is key to
enhance productivity. To accomplish given objectives and achieve the desired targets, three
major approaches for yield improvement in any crop may be pursued by way of:
Evaluation of variation within existing crop cultivars and selecting promising
lines/genotypes.
Development of variable material through genetic recombination or other means
and selecting the most promising lines for further manipulation.
Transformation of gene/genes for quantitative traits from wild relatives to the
cultivated species either through conventional breeding techniques or through genetic
engineering.
The breeding procedure for yield and yield related traits depends upon the pattern of
inheritance (qualitative or quantitative), the number of genes with relative effects, and the
nature of these genes. There is now considerable body of information available about the
variation both between and within crop species in response to yield and its related traits.
The knowledge of genetic control of the characters is important in organizing any
kind of plant breeding program. Information regarding the genetic architecture of
economically important characters, number of genes controlling each trait, heritability
estimates and genetic correlations between yield and yield components are essential to
exploit variation in the breeding material. Genetic analysis of parents and the F1 alone
provides satisfactory assessment of genetical control of a character.
3
The present study pertaining to genetic analysis of seed yield and its contributing
components and inheritance of wilt resistance was planned with the following objectives.
1. To estimate the genetic behavior i.e. gene action for seed yield and its
components.
2. To work out general and specific combining ability mean squares, variances and
their effects.
3. To find difference among the parents and their F1 by using DNA finger printing.
4. To find the nature of inheritance of Fusarium oxysporum, sp. Ciceris resistance
using chi-square test.
4
Chapter- 2
REVIEW OF LITERATURE
1. Gene action studies
Selection of desired plant type possessing genes for high yield and disease
resistance is effective in situations when the traits are controlled by predominantly additive
gene action in chickpea genotypes. If a trait is expressed through over dominance type of
gene action, it is due to the presence of interactions of non-additive nature. The type of
gene action for the expression of a certain trait depends upon the gene frequency in the
genotypes used as parents. Recombination’s shows additive or non-additive type of gene
action for the expression of a trait. Variations in the mode of inheritance for various traits
in chickpea has been reported in the literature and presented as below.
Number of days taken to flowering
Mode of inheritance for number of days taken to flowering has been variably
described in the literature however in majority of the cases additive genetic variation has
been reported for this trait in chickpea genotypes. Sandhu and Mandal (1989), Singh et al.
(1992), Anbessa et al. (2006), Bicer and Sakar (2008) reported number of days taken to
flowering predominantly under the control of additive type of gene action in chickpea
genotypes. Slightly different findings were published by Patil et al. (1998) and Biranvand
(2008) who reported additive and dominance type of gene action for number of days taken
to flowering in chickpea genotypes. Kulkarni (2001), Sarode et al. (2001), Patil et al.
(2006), Meena et al. (2006) and Patil et al. (2006) reported non-additive type of gene
action for this trait. Derya et al. (2006) detected 5.47% broad sense heritability. Durga et
al. (2007) observed additive gene effects with high heritability for this trait. Saleem et al.
(2005), Arshad et al. (2004) and Sidramappa et al. (2008) reported high heritability with
low genetic advance for number of days taken to flowering in chickpea genotypes. Deb and
Khaleque (2009) observed low heritability both in narrow and broad senses for number of
days taken to flowering in chickpea genotypes.
5
Number of Primary Branches per Plant
Mode of inheritance for number of primary branches per plant has been variably
described in literature and most of the studies revealed that additive and non- additive type
of gene action for this trait in chickpea genotypes. Singh et al. (1982), Malhotra et al.
(1983) and Yadav et al. (1987) observed additive type of gene action for this trait in
chickpea genotypes. In contrast Meena et al. (2006) observed non-additive gene action for
this trait in chickpea genotypes. Singh and Bains (1982), Singh et al. (1992), Path et al.
(1998), Bicer and Sakar (2008) reported additive and non-additive type of gene action for
number of primary branches per plant in chickpea genotypes. Deb and Khaleque (2009)
observed low heritability both in narrow and broad senses for number of primary branches
per plant in chickpea genotypes.
Number of Secondary Branches per Plant
There are variable findings pertaining to the genetic control of number of secondary
branches per plant in chickpea genotypes. Singh et al. (1992), Sarode et al. (2001), Bicer
and Sakar (2008) and Sreelakshmi et al. (2010) reported additive and dominant gene effects
for the trait. Slightly variable results have been reported by Singh et al. (1982), Malhotra
et al. (1983), Yadav et al. (1987), Kulkarni (2001), Arshad et al. (2004) and Patil et al.
(2006) who indicated additive type of gene action for number of secondary branches per
plant. Singh and Bains (1982), Tewari and Pandey (1986) and Patil et al. (1998) observed
predominance of additive gene action but non-additive effects were also significant. In
contrast Meena et al. (2006) reported non-additive type of gene action for this trait. Girase
and Deshmukh (2000) observed duplicate type of gene action for the expression of number
of secondary branches per plant in chickpea genotypes. Deb and Khaleque (2009) observed
low heritability both in narrow and broad senses for number of secondary branches per
plant in chickpea genotypes.
6
Plant Height (cm)
Singh et al (1991), Kulkarni (2001), Patil et al. (2006),Bicer and Sakar (2008)
reported significant additive gene action for plant height in chickpea genotypes. Slightly
variable findings were published by Beyran et al. (2008) and Singh et al (1992) who
detected additive and dominance gene action for this trait. In contrast Sarode et al. (2001)
and Meena et al. (2006) observed non-additive type of gene action. Biranvand et al. (2008)
estimated genetic parameters D, H1 and H2 and indicated that additive variance component
was greater than non-additive component, narrow sense heritability was medium and
Wr/Vr graphs reflected partial dominance for plant height in chickpea genotypes.
Sidramappa et al. (2008) observed high estimates of heritability for plant height in
chickpea genotypes.
Number of days taken to maturity
Singh et al.(1992), Bicer and Sakar (2008) and Biranvand (2008) reported additive,
dominant gene effects for this trait in chickpea genotypes. In contrast Sarode et al. (2001)
and Meena et al. (2006) observed non-additive type of gene action in chickpea genotypes.
Arshad et al. (2004) detected high heritability with low genetic advance for this trait.
Girase and Deshmukh (2000) observed duplicate type of gene action for the expression of
days taken to maturity in chickpea genotypes.
Total weight of plant (g)
The literature revealed total weight of plant to be under the control of both additive
and non-additive type of gene action in chickpea genotypes. Singh and Bains (1985),
Tewari and Pandey (1986) and Arshad et al. (2004) reported additive gene effects to be
important. In contrast Meena et al. (2006) observed non-additive type of gene action for
this trait in chickpea genotypes. Singh and Bains (1982) observed that both additive and
non-additive gene action were important for total weight of plant. Hussain et al. (1990)
found over-dominant type of gene action for total plant weight. Venkatraman (2007)
indicated the predominant role of additive gene action for biomass in chickpea genotypes.
7
Number of pods per plant
A wide range of literature is available on the inheritance pattern of number of pods
per plant in chickpea genotypes. Singh et al. (1982), Tewari and Pandey (1986), Yadav et
al. (1987), Singh et al (1991), Durga et al. (2007) Bicer and Sakar (2008) and Sreelakshmi
et al. (2010) reported significant additive gene action in chickpea genotypes. Slightly
variable results have been reported by Singh et al. (1992), Chaturvedi et al. (1997) and
Beyran et al. (2008) also found additive and dominance gene action. In contrast Kulkarni
(2001), Sarode et al. (2001), Meena et al (2006) and Patil et al. (2006) reported non-
additive type of gene action. Kumar et al. (2001) revealed the predominance of non-
additive variance for this trait. Singh and Bains (1982) and Malhotra et al. (1983) observed
both additive and non-additive gene action to be important with additive prevailing.
Hussain et al. (1990) found that mode of inheritance to be under the control of over-
dominant gene action for number of pods per plant in chickpea genotypes. Patil et al.
(1998) observed preponderance of epistatic gene action for number of pods per plant.
Annigeri et al. (1996) revealed the predominance of additive variance for pods per plant in
chickpea genotypes. Biranvand et al. (2008) estimated genetic parameters D, H1 and H2
and indicated that additive variance component was less than non-additive variance
components, narrow sense heritability was low and Wr/Vr graphs reflected over dominance
for number of pods per plant. Sidramappa et al. (2008) observed high estimates of
heritability for pods per plant in chickpea genotypes.
Number of seeds per pod
Mode of inheritance for number of seeds per pod in chickpea genotypes has been
described in literature. Chaturvedi et al. (1997), Sarode et al. (2001) reported additive and
dominance gene action. Yadav et al. (1987) Singh et al. (1992) and Beyran et al. (2008)
detected additive gene action for this trait in chickpea genotypes. Biranvand et al. (2008)
estimated genetic parameters D, H1 and H2 which indicated that additive variance
component was less than non-additive variance component, narrow sense heritability was
low and Wr/Vr graphs reflected over dominance for number of seed per pod. Singh and
Bains (1982) and Malhotra et al. (1983) observed both additive and non-additive gene
action were important with additive predominating. However, non-additive type of gene
8
action has been reported by Meena et al. (2006). Martinez et al. (1978) observed over-
dominance in a positive sense for seeds per pod. Patil et al. (1998) observed predominance
of epistatic gene action. Sandhu and Mandal (1989) and Annigeri et al. (1996) revealed the
predominance of additive variance for seeds per pod. Kumar et al. (2001) revealed the
predominance of non-additive variance for this trait in chickpea genotypes. Deb and
Khaleque (2009) observed low heritability both in narrow and broad senses for seeds per
pod in chickpea genotypes.
100-seed weight (g)
100-seed weight is an important yield contributing trait in chickpea genotypes.
Chaturvedi et al. (1997), Bicer and Sakar (2008) reported significant additive, dominant
gene effects, high narrow-sense heritability (96%) and reciprocal differences for this trait.
Singh et al. (1982), Singh et al. (1992), Kulkarni (2001) and Patil et al. (2006) reported
additive type of gene action in chickpea genotypes. Sandhu and Mandal (1989) and Kumar
et al. (2001) revealed the predominance of additive variance for 100-seed weight. Singh
and Bains (1982), Sarode et al. (2001) and Meena et al. (2006) detected non-additive type
of gene action. Malhotra et al. (1983), Annigeri et al. (1996) revealed both additive and
non-additive variance for this trait. Saleem et al. (2005) and Arshad et al. (2004) and
Sidramappa et al. (2008) observed high heritability with low genetic advance for this trait.
Girase and Deshmukh (2000) observed duplicate type of gene action for the expression of
100-seed weight in chickpea genotypes. Deb and Khaleque (2009) observed low
heritability both in narrow and broad senses for 100-seed weight in chickpea genotypes.
Grain yield per plant (g)
Grain yield per plant in chickpea genotypes is a complex trait controlled by multiple
genes. Grain yield per plant has been reported under the control of additive and dominance,
non-additive, dominance, over dominance, additive and partially additive effects of genes
in chickpea genotypes. Singh et al. (1992), Chaturvedi et al. (1997), Sarode et al. (2001)
and Beyran et al. (2008) and Sreelakshmi et al. (2010) reported additive and dominance
gene action. Singh and Bains (1982), Malhotra et al. (1983), Tewari and Pandey (1986) and
Annigeri et al. (1996) observed both additive and non-additive genetic variance to be
9
significant for this trait. Yadav et al. (1987) and Durga et al. (2007) reported additive gene
effects with high heritability. Kulkarni (2001), Meena et al. (2006) and Patil et al. (2006)
observed non-additive type of gene action for this trait in chickpea genotypes. Patil et al.
(1998) observed predominance of epistatic gene action. Kumar et al. (2001) revealed the
predominance of non-additive variance for this trait. Husain et al. (1990) found that mode
of inheritance appeared to be over-dominant for grain yield per plant. Martinez et al.
(1978) observed over-dominance in a positive sense and weak reciprocal differences for
seeds per pod in chickpea genotypes. Biranvand et al. (2008) estimated genetic parameters
D, H1 and H2, indicated that additive variance component was less than non-additive
variance component, narrow sense heritability was low and Wr/Vr graphs reflected over
dominance for seed yield per plant. Girase and Deshmukh (2000) observed grain yield to
be predominantly under the control of dominance effect. Sidramappa et al. (2008) observed
high estimates of heritability for seed yield per plant in chickpea genotypes. Deb and
Khaleque (2009) observed low heritability both in narrow and broad senses for grain yield
per plant in chickpea genotypes.
Harvest index %
Venkatraman (2007) indicated predominant role of additive gene action for harvest
index in chickpea genotypes. In contrast Sandhu and Mandal (1989), Kulkarni (2001) and
Patil et al. (2006) observed non-additive type of gene action for this trait. Chaturvedi et al.
(1997) reported additive and non-additive gene action for harvest index in chickpea
genotypes.
2. Combining ability studies
Number of days taken to flowering
Singh et al. (1992) found additive gene action to be predominant and highly
predictable for this trait in chickpea genotypes. Both general combining ability (GCA) and
specific combining ability (SCA) effects varied significantly. Anbessa et al. (2006), Bicer
and Sakar (2008) and Bhardwaj et al. (2009) revealed that additive variance was significant
in chickpea genotypes. Kulkarni (2001) observed that mean sum of squares due to general
and specific combining ability was highly significant in chickpea genotypes. Patil et al.
(2006) reported variances due to general and specific combining abilities to be significant.
10
JG1265, K850, Phule G12 and BDN9-3 were good combining parents for early or late
flowering in chickpea genotypes (Sandhu and Mandal, 1989).
Number of days taken to maturity
Number of days taken to maturity is an important yield related trait and plant
breeders are always interested in developing short-duration varieties. Kabeta (2006),
Anbessa et al. (2006) Bicer and Sakar (2008) revealed that additive variance was
significant in chickpea genotypes. Patil et al. (2006) reported variances due to general and
specific combining abilities to be non-significant. Additive, dominant gene effects have
been reported by Singh et al. (1992), Bicer and Sakar (2008) and Biranvand (2008). In
contrast Meena et al (2006) observed non-additive type of gene action for the inheritance
of this trait in chickpea genotypes.
Plant Height
Katiyar et al.(1988) observed significant role of additive and non-additive gene
action, with former being predominant for days taken to flowering in chickpea genotypes.
Bicer and Sakar (2008) and Bhardwaj et al. (2009) revealed that additive variance was
significant. Kulkarni (2001) observed mean sum of squares due to general and specific
combining ability as highly significant. Patil et al. (2006) reported variances due to general
and specific combining abilities to be significant in chickpea genotypes.
Number of Primary Branches per Plant
Different type of gene action for number of primary branches per plant has been
described in the literature however, in most cases both additive and non-additive type of
gene action reported for this trait in chickpea genotypes. Katiyar et al. (1988) revealed
significant role of additive and non-additive gene action, with former being predominant
for number of primary branches per plant in chickpea genotypes. Malhotra et al. (1983),
Yadav et al. (1987), Bicer and Sakar (2008) and Bhardwaj et al. (2009) reported additive
variance to be significant. Kulkarni (2001) observed mean sum of squares due to general
and specific combining ability highly significant in chickpea genotypes.
11
Number of Secondary Branches per Plant
Tewari and Pandey (1986) and Katiyar et al.(1988) revealed significant role of
additive and non-additive gene actions, with former being predominant for number of
secondary branches per plant in chickpea genotypes. Malhotra et al. (1983), Yadav et al.
(1987), Bicer and Sakar (2008) and Bhardwaj et al. (2009) reported additive variance to be
significant in chickpea genotypes. Kulkarni (2001) observed mean sum of squares due to
general and specific combining ability to be highly significant. Patil et al. (2006) reported
variances due to general and specific combining abilities significant in chickpea genotypes.
Number of pods per plant
A wide range of literature is available on the inheritance pattern of number of pods
per plant in chickpea genotypes. Malhotra et al. (1983), Tewari and Pandey (1986) and
Katiyar et al. (1988) revealed significant additive and non-additive gene actions for this
trait. Yadav et al. (1987), Bicer and Sakar (2008) revealed that additive variance was
significant. Hussain et al. (1990) assessed general and specific combining abilities and
found that mode of inheritance appeared to be over-dominant for this trait. Kulkarni (2001)
observed mean sum of squares due to general and specific combining ability were highly
significant in chickpea genotypes. Patil et al. (2006) reported variances due to general and
specific combining abilities significant. Chaturvedi (1997) revealed both additive as well as
non-additive components of variance to be important in the expression of this trait and per
se performance may be used as an index of GCA effects in chickpea genotypes.
Number of seeds per pod
Mode of inheritance for number of seeds per pod has been variably described in
literature in chickpea genotypes. Bicer and Sakar (2008) and Bhardwaj et al. (2009)
reported additive variance as significant in chickpea genotypes. Kulkarni (2001) observed
highly significant mean sum of squares due to general and specific combining ability.
Malhotra et al. (1983) and Chaturvedi (1997) revealed both additive as well as non-additive
components of variance to be important in the expression for number of seeds per pod in
chickpea genotypes.
12
Total weight of plant
The literature revealed for total weight of plant to be under the control of both
additive and non-additive type of gene action in chickpea genotypes. Bicer and Sakar
(2008) and Bhardwaj et al. (2009) reported significant additive variance for this trait.
Kulkarni (2001) observed highly significant mean sum of squares due to general and
specific combining ability. Venkatraman (2007) observed GCA variances significant and of
a high magnitude, while specific combining ability (SCA) variances were non-significant,
indicating the predominant role of additive gene action on the biomass. Tewari and Pandey
(1986) revealed significant role of additive and non-additive gene actions. Hussain et al.
(1990) assessed general and specific combining abilities and found that mode of
inheritance appeared to be over-dominant for this trait. Singh and Bains (1985) and Arshad
et al. (2004) indicated additive gene effects to be important in chickpea genotypes. Meena
et al. (2006) reported non-additive type of gene action for this trait in chickpea genotypes
Venkatraman (2007) estimated GCA variances were significant and of a high magnitude,
while specific combining ability (SCA) variances were non-significant, indicating the
predominant role of additive gene action on biomass in chickpea genotypes.
100-seed weight (g)
100-seed weight is an important yield contributing trait in chickpea genotypes.
Genetics for chickpea genotypes has been described in the literature. Katiyar et al. (1988)
revealed significant role of additive and non-additive gene actions, with former being
predominant for 100-seed weight in chickpea genotypes. Malhotra et al. (1983) observed
both additive and non-additive gene action were important with additive predominating in
chickpea genotypes. Bicer and Sakar (2008) and Bhardwaj et al. (2009) revealed that
additive variance was significant. Kulkarni (2001) observed mean sum of squares due to
general and specific combining ability as highly significant. Patil et al. (2006) reported
Variances due to general and specific combining abilities as significant. Chaturvedi (1997)
revealed both additive as well as non-additive components of variance important in the
expression of this trait. Sandhu and Mandal (1989) revealed that the per se performance
may be used as an index of GCA effects. JG1265, K850, Phule G12 and BDN9-3 were
good combining parents for seed weight in chickpea genotypes.
13
Grain yield per plant (g) Grain yield per plant is complex character and complex is its inheritance. Tewari
and Pandey (1986) and Katiyar et al. (1988) revealed significant role of additive and non-
additive gene actions, with former being predominant for days taken to flowering in
chickpea genotypes. Malhotra et al. (1983) observed both additive and non-additive gene
action important with additive predominating. Kulkarni (2001) observed mean sum of
squares due to general and specific combining ability as highly significant. The best cross
combinations involved parents with high and low gca effects. Yadav et al. (1987) and
Bhardwaj et al. (2009) revealed that additive variance was significant. Patil et al. (2006)
reported variances due to general and specific combining abilities as significant.
Chaturvedi (1997) revealed both additive as well as non-additive components of variance
were found important in the expression of this trait in chickpea genotypes. The study
revealed that the per se performance may be used as an index of GCA effects. Hussain et
al. (1990) assessed general and specific combining abilities and found that mode of
inheritance appeared to be over-dominant for grain yield per plant in chickpea genotypes.
Harvest index %
Venkatraman (2007) observed GCA variances significant and of a high magnitude,
while specific combining ability (SCA) variances were non-significant, indicating the
predominant role of additive gene action on harvest index in chickpea genotypes. Kulkarni
(2001) observed mean sum of squares due to general and specific combining ability as
highly significant. Patil et al. (2006) reported variances due to general and specific
combining abilities as significant. Bhardwaj et al. (2009) revealded additive and
Chaturvedi (1997) revealed both additive as well as non-additive components of variance
important in the expression of this trait. Sandhu and Mandal (1989) revealed that the per se
performance may be used as an index of GCA effects. JG1265, K850, Phule G12 and BDN9-3
were good combining parents for harvest index in chickpea genotypes.
14
Epigenetics
Epigenetics refers to DNA and chromatin modifications that persist from one cell
division to the next despite a lack of change in the underlying DNA sequence. Some
epigenetic changes show transgenerational inheritance meaning that these changes can be
passed from one generation to the next. Epigenetics plays an important role in cellular
differentiation, allowing dinstict cell types to have specific characteristics despite sharing the
same DNA sequence. Some examples of epigenetic processes include imprinting,
bookmarking, gene silencing, paramutation, X chromosome inactivation, reprogramming,
position effect, histone modifications and heterochromatin, some carcinogenesis, maternal
effects, and transvection. The epigenome refers to the overall epigenetic state of a cell while
the epigenetic code refers to epigenetic features such as DNA methylation and histone
modifications that create different phenotypes in different cells.
Epigenetic mechanisms
The mechanisms of epigenetic inheritance systems are still being unraveled, but our
current understanding has uncovered at least 4 routes by which epigenetic changes persist
over time. These routes include RNA transcripts, cellular structures, DNA
methylation/chromatin remodeling. The expression of certain genes can enhance the
production of other similar genes, and even the gene itself. The enhanced expression of
certain genes can be inherited by subsequent descendents. This can be through transcription
factors and signal transduction pathways; in systems containing gap junctions, the actual
mRNA may be spread to other cells or nuclei via diffusion.
Importance of epigenetics
Epigenetics is important in development partly because the genome is largely static.
Epigenetic modifications are required to provide some dynamic variability to cellular
function and phenotype, allowing for cellular differentiation. Epigenetics may also be
important in evolution since transgenerational inheritance of epigenetic modifications has
been observed (e.g., paramutation in maize). Often however, these modifications are lost
after several generations.
15
3. Molecular Studies
Welsh and McCielland (1990) described that simple and reproducible fingerprinting
of complex genomes could be generated using single arbitrarily chosen primers and
polymerase chain reaction (PCR). No prior sequence information was required. Their
method, arbitrarily primed PCR, involve two cycles of low stringency amplification
followed by PCR at higher stringency. They showed that strains can be distinguished by
comparing polymorphisms in genomic fingerprints. The generality of the method was
demonstrated by application to 24 strains from five species of Staphylococcus, eleven
strains of Staphylococcus pyogenes and three varieties of Oryza sativa. Williams et al.
(1990) studied random primers in PCR to identify DNA markers linked to the trait of
interest. The polymorphisms were inherited in a simple Mendelian fashion and they called
these polymorphisms as RAPD markers. They suggested that these markers could be used
for DNA fingerprinting.
Rakesh et al. (2002) used PCR-based RAPD markers to assess diversity in 23
chickpea genotypes. 40 of 100 random primers had shown polymorphism. Most of the
primers revealed single band and only 14 were polymorphic. It was observed that RAPD
analysis employing 30 polymorphic primers could provide better estimates of genetic
relationship in chickpea. Souframanien and Gopalakrishna (2004) used RAPD to study the
DNA polymorphism in elite blackgram genotypes. A total of 25 random primers were used.
Amplification of genomic DNA of the 18 genotypes, using RAPD analysis, yielded 104
fragments that could be scored, of which 44 were polymorphic, with an average of 1.8
polymorphic fragments per primer. Cingilli and Akcin (2005) studied genetic diversity in
chickpea using a simple and reliable DNA extraction method. This small scale method is
CTAB based and extract DNA from 1 to 3 folded young leaves processed in a 1.5 ml tube
with 0.5 ml of extraction buffer and homogenized. High quality DNA was obtained and
used for chain reaction amplifications using the mini-prep CTAB method. Rao et al.
(2007) premeditated genetic relationships between 19 chickpea cultivars and five
accessions of its wild progenitor by using RAPD markers. On an average, six bands per
primer were observed in RAPD analysis. In RAPD analysis 51.7% and 50.5% polymorphic
bands were observed among wild accessions and chickpea cultivars, respectively. The
16
markers generated by RAPD assays can provide practical information for the management
of genetic resources.
Talebi et al. (2009) studied genetic variation among cultivated chickpea and six
other related wild species of Cicer with RAPD. Among the 42 random 10-mer primers
tested, only 9 amplified genomic DNA across all species. Three main species groups were
identified by UPGMA clustering using Li’s pair-wise calculations. Khan et al. (2010) used
RAPD analysis for studies of genetic diversity of cotton genotypes. Out of 45 decamer
randomly sequenced primers applied for RAPD analysis, 25 showed polymorphism. All of
25 primers produced 205 fragments, out of which 144 were polymorphic accounting for
70.24% of the total number of fragments. The study demonstrated the efficient and reliable
use of RAPDs for analyzing genetic variation in coloured and white-linted genotypes of
cotton.
3. Wilt inheritance studies
Pathak et al. (1975), Kumar and Haware (1982), Upadhyaya et al. (1983) and
Kumar (1998) studied differences in time of wilting of chickpea (Cicer arietinum L.) in
response to Race 1 of Fusarium oxysporum f.sp. ciceris. C-104 wilts later than JG-62 and
the difference in time of wilting appears to be inherited as a single gene with early wilting
partially dominant to late wilting. Considered in relation to earlier studies, the observations
indicate that at least two genes are involved in the inheritance of resistance in chickpea to
Race 1 and offer an explanation for previous difficulties in interpreting the inheritance of
resistance. Farooq et al. (2005) and Pande et al. (2007) conducted physiological studies of
Fusarium oxysporum F. sp. ciceri.
Gumber et al. (1995) indicated that the resistance to race 2 of Fusarium wilt is
controlled by two genes, the first of which must be present in the homozygous recessive
form, and the other in the dominant form, whether homozygous or heterozygous for
complete resistance. Early wilting results if the other gene is homozygous recessive. Late
wilting occurs if both loci are dominant. The existence of differences among chickpea
cultivars in the time taken to express the initial symptoms of Fusarium wilt were observed.
Navas-Cortés et al. (2000) studied the losses due to development of Fusarium wilt
epidemics. Shah et al. (2009) screened two hundred and forty nine chickpea mutants
developed through gamma irradiation along with their respective parents and susceptible
17
check AUG-424 for resistance to Fusarium wilt in natural wilt sick plot during 2003-2004
seasons. All the 4 parent genotypes showed highly susceptible reaction to Fusarium wilt.
Out of a total of 249 morphological mutants of 4 genotypes, 75 mutants exhibited highly
resistant reaction (less than 10 %) followed by 31 mutants resistant (11 to 20%), 34
mutants moderately resistant / tolerant (21 to 30%), 35 mutants susceptible (31 to 50%)
and 75 mutants were highly susceptible (50 to100%).
Ahmad et al. (2010) indicated that wilt caused by Fusarium oxysporum
Schlechtend.Fr. f. sp. ciceris is a devastating disease of chickpea in Pakistan. 321
genotypes from different sources were evaluated under controlled condition to identify
genetic sources of resistance against this disease at seedling and reproductive stage.
Disease reaction at two stages revealed considerable variation among the genotypes. At
seedling stage disease incidence varied from 0 to 29.3% whereas at reproductive stage
ranged from 0 to 57%. At seedling stage 173 genotypes were resistant, 54 were tolerant and
94 were susceptible, whereas at reproductive stage, 102 genotypes were resistant, 36 were
tolerant and 183 were susceptible. Eighty two genotypes showed steady resistance at both
stages. These genotypes may be exploited for the development of resistant cultivars against
wilt.
18
Chapter-3
MATERIALS AND METHODS
The present study was carried out in the experimental area of the Department of
Plant Breeding and Genetics, University of Agriculture, Faisalabad during the years 2006-
2009. The experimental material comprised six diverse chickpea genotypes i.e., CM-98,
AUG-786, Bittal-98, Balksar-2000, Wanhar-2000 and Punjab-2000 homozygous in nature,
maintained by the Department. The data were recorded these parental lines (Table.1). The
parental lines exhibited a range of variation for seed yield and other traits. These genotypes
were sown in the field during the year 2006-2007 and crossed in all possible combinations.
Chickpea is a highly self pollinated plant, the stigma becomes receptive about 12 hours
before anther dehiscence which helps in making controlled pollinations. Emasculation was
done in late afternoon one day prior to pollination. Emasculation was done by opening the
sepals enclosing the keel, opening keel and removing the stamens by firmly grasping the
filaments. The emasculated flowers were covered and tagged for identification. Pollination
was made following day in early morning between 8 to 9 a.m. A large number of crosses
were attempted. At maturity the seed was harvested from the female parents.
Assessment of F1 and Parental Lines
During the year 2007-2008, all possible F1 crosses along with their parents were
sown in a randomized complete block design with three replications. Each treatment
consisted of single row with row to row and plant to plant spacing of 30 cm and 15 cm,
respectively. Two seeds per hill were sown by using dibbled which were thinned later to a
single healthy seedling per hill. All field operations including hoeing, weeding etc were
carried out uniformly to reduce experimental error. Except for days taken flowering and
number of days to maturity data on ten equally competitive plants from each treatment
were recorded at maturity for the following quantitative traits.
19
Table.1. Characteristics of parental genotypes
Trait CM-98 Bittal- 98
AUG-786
Punjab-2000
Wanhar-2000
Balksar-2000
Number of days taken
to flowering
120.3 113.0 117.7 115.3 113.0 123.0
Number of primary
branches per plant
2.66 2.57 2.23 2.94 2.46 2.19
Number of secondary
branches per plant
6.63 5.76 5.13 6.81 5.55 5.43
Plant height (cm) 68.0 62.7 74.3 56.0 62.7 58.7
Number of days taken
to maturity
161.3 169.7 160.3 153.3 163.7 154.0
Total weight of plant
(g)
85.9 82.2 84.3 77.9 72.5 66.4
Number of pods per
plant
94.0 89.7 90.0 84.0 75.0 87.7
Number of seeds per
pod
1.74 1.75 1.72 1.71 1.82 1.73
100-seed weight (g) 23.61 23.68 23.52 22.24 21.34 20.13
Grain yield per plant
(g)
40.8 37.2 36.3 37.0 29.1 31.3
Harvest index % 47.5 45.3 43.0 40.9 40.1 47.2
20
Number of days taken to flowering
Data for number of days taken to flowering were recorded as the number of days
from the date of planting to the time when 50 % plants within a plot showed the
appearance of first flower. The average numbers of days taken to flowering were
calculated.
Number of primary branches per plant
The branches emerging from the crown (base at ground level) of the plant were
considered as primary branches. The total number of primary branches per plant were
counted at the time of the harvest and averaged.
Number of secondary branches per plant
The branches emerging from the primary branches were counted in each of the
selected plant as the number of secondary branches and then averaged.
Plant height (cm)
Height of the earmarked plants was measured in centimeters from the ground level
to the canopy top with the help of a meter rod when no more growth was expected and
averaged.
Days taken to maturity
The data were recorded at the time of maturity when 90% plants of a plot turned
brown and ready for harvest. Number of days were calculated from the date of planting to
the date of maturity and averaged.
Total weight of plant (g)
Dried plants were harvested carefully without losing any debris and kept in kraft
paper bags at room temperature. Their weight was recorded in grams by using electronic
balance after 15 days of the harvest.
Number of pods per plant
Every possible care was taken to avoid the shedding of pods at the time of harvest.
Immediately after harvest, the pods from each plant were removed, counted and kept in
separate craft paper bag properly labeled. The average number of pods per plant was
computed.
21
Number of seeds per pod
Ten pods were randomly selected from each plant. These were threshed manually
and seeds were counted and average numbers of seeds per pod were calculated.
100-Seed weight (g)
For the calculation of seed weight 100 seeds from the bulk produce of the selected
plants of each entry was taken and weighed in grams by using an electronic balance and the
average was worked out.
Grain yield per plant (g)
Total quantity of threshed seeds obtained from each of the selected plant was
collected in the kraft paper bag. The grain yield of each plant was weighed in grams with
the help of an electronic balance. The average was obtained by dividing the grain yield of
the selected plants by their number.
Harvest index (%)
Harvest index was calculated by using the following formula:
Harvest index = _ Grain yield per plant × 100
Biomass per plant
Statistical Analysis
Analysis of Variance
Before subjecting the data to diallel analysis an ordinary analysis of variance (Steel
and Torrie, 1994) was performed to determine whether the genotypic differences were
significant for the characters under consideration.
1. Genetic Analysis
Genetic analysis following additive-dominance model was done using a diallel
mating scheme as described by Hayman (1954a, b, 1958) and Jinks (1954).
Assumptions of Diallel Analysis and Test for their variance
Diallel analysis is based on six assumptions as described by Hayman (1954b). These
include:
o Normal diploid segregation
o Absence of reciprocal effects
o Homozygous parents
22
o Absence of epistasis
o No multiple allelism
o Independent gene distribution
Analysis of the diallel table was carried out to obtain preliminary information about
the presence of additive (a), dominance (b), maternal (c) and reciprocal (d) effects. The b
item was further partitioned into b1, b2 and b3 items. The b1 tests directional dominance
effect, b2 tests effects due to parents contributing varying degree of dominance alleles and
b3 tests that part of the dominance deviation that is unique to each F1 (specific gene
interaction). On the assumption of no genotype × environment" interaction and no
differences between reciprocal crosses the mean squares for c, d and block interactions are
all estimates of E, the environmental component of variation. If reciprocal crosses differ, c
detects the average maternal effects of each parental line and the d reciprocal differences
not ascribable to c. Any genotype × environment interactions are detected as a difference
between the block interactions for the a and b items indicating that additive and dominance
variations were influenced to a greater extent by the environment.
Information about gene action was inferred by plotting the covariance (Wr) of each
array against its variance (Vr). The slope and position of the regression line fitted to the
array points within the limiting parabola indicated the degree of dominance and the
presence or absence of gene interaction.
The points for limiting parabola (Wri) were obtained as follows:
ii rr VVpW
Where, Vp = parental variance, and Vri = array variance.
Array variances and co-variances were used to draw a regression line within the
limiting parabola. The distance between the origin and the point where the regression line
cut the Wr-axis provides measure of average degree of dominance:
i. Partial dominance: when the intercept is positive
ii. Over dominance: when the intercept is negative, and
iii. No dominance: when the regression line touches the parabola limits.
The order of array points represents the distribution of dominant and recessive genes
among the parents. The parents with most dominant genes fall nearest to the origin while
the parents with most recessive genes fall farthest from the origin, and the parents with
23
equal frequencies of dominant and recessive genes fall in the middle. The standard error of
the regression line slope was estimated according to Johnson and Askel (1964).
Chickpea (Cicer arietinum L.) is a diploid species having 8 haploid chromosome
numbers and therefore behaves cytogenetically as a diploid species. For removing
reciprocal differences the values in the off-diagonal cells of the diallel table are replaced by
means of the direct and reciprocal cross prior to analysis. The parental genotypes included
in the study were selected from the gene pool of University of Agriculture Faisalabad
maintained every year.
To determine the assumptions of the absence of epistasis, no multiple allelism and
independent gene distribution, the data were subjected to two tests. The first test was a joint
regression analysis of Vr (variance of each array) and Wr (Parent-offspring covariance),
estimated from the mean diallel table. Then the regression of covariance on the variances
was computed. According to Mather and Jinks (1982) the regression coefficient is expected
to be significantly different from zero but not from unity. Failure of this test means that
non-allelic interaction (epistasis) is present or genes are not independent in their action, or
show non-random association among the parents.
The second test for the adequacy of additive-dominance model is the analysis of
variance of the Wr+Vr and Wr-Vr. If dominance (or certain types of non-allelic interaction)
is present, Wr + Vr must change from array to array. Similarly if epistasis exists, Wr-Vr
will vary between arrays.
Failure of both the tests completely invalidates the additive-dominance model.
However, if one of them fulfills the assumption, the additive-dominance model was
considered partially adequate. Components of variance for such type of partially adequate
models have also been estimated (Johnson and Askel, 1964; Wilson et al., 1978).
24
Genetic Components of Variation
The genetic components of variation were calculated using the procedures given by
Hayman (1954a) and Mather and Jinks (1982). The genetic parameters were estimated
as follows:
Additive variation (D)
EVD p
Where
Vp = varianceof the parents
E = environmental variance
Variation due to dominant effect of genes (H1)
En
nVrWrVH p
23441
Where
Vr = mean of the array variances
Wr= mean of the covariance’s between parents and arrays
n = number of parents.
Variation due to dominant effect of genes correlated for gene
distribution (H2)
EVmVrH 2442
Where
Vm = variance of the mean of arrays.
Relative frequency of dominant and recessive alleles (F) In the presence of unequal gene frequencies, the sign and magnitude of F
determines the relative frequency of dominant and recessive alleles in the parental
population and the variation in the dominance level loci. F is positive whenever the
dominant alleles are more than the recessive ones, irrespective of whether these are
increasing or decreasing in their effect. It was calculated as follows:
E
n
nWrVF p
2242
25
Overall dominance effect of heterozygous loci
E
n
nMMh LOLI 2
2 144
Where
2
2 ./.1
valueparentsnTGn
MM LOLI
26
Environmental variance (E)
pdfErrordf
pSSErrorSSE
Re
Re/No. of Reps
Where,
Error SS = error sum of square
Rep. SS = replication sum of squares in the analysis of variance.
Average degree of dominance
DH /
Proportion of genes with positive and negative effects in the
parents
uv over all loci. u = frequency of increasing alleles, and v = 1-u
.frequency of decreasing alleles. This is equal to 0.25 when n = v at all loci
(Singh and Chaudhary, 1985).
12 4/ HH
Proportion of dominant and recessive genes
FDH
FDH
14
14
Heritability (h2)
Broad Sense h2 (bs) =EFHHD
FHHD
5.0225.015.05.0
5.025.015.05.0
Narrow Sense h2 (ns) =EFHHD
FHHD
5.0225.015.05.0
5.0225.015.05.0
2. Combining Ability Analysis
Assuming no differences among the direct and reciprocal crosses, the mean
performance of a cross (XAB) should be equal to GCAA + GCAB + SCAAB. The GCAA and
GCAB is the general combining ability of the A and B parents and performance of a cross
of A and B is expected to be equal to the sum (GCAA + GCAB) of general combining
27
ability of parents, However, the actual performance of the cross may be different from this,
sum by an amount equal to specific combining ability (SCA). In terms of gene action, the
differences in GCA are due to additive genetic variance and additive x additive type of
epistasis, where as SCA estimates non-additive genetic variance.
Following the concept of GCA and SCA (Sprague and Tatum, 1942), several
methods of analysis were developed. Griffing (1956) provided detailed procedures for the
analysis of variance and estimation of combining ability effects under two models (Model
1-fixed; Model 11-random), and four methods.Method-I: Parents, one set of Fls and
reciprocal Fls are included (n2) Method -II: Parents and Fls are included, but reciprocal are
not (1/2) n (n-1) Method-III: One set of F1s and reciprocal are included but not the
parents(n)(n-1) Method-IV; One set of Fls but neither parents nor reciprocal FlS is included
(1/2 (n (n-1)
The method-I model-II of Griffing (1956) was applied here for combining ability
analysis of the data.
3. Molecular Study
(a) Assessment of genetic diversity among parents and their F1s using
RAPD.
With the advent of polymerase chain reaction (PCR) technology, it has become
possible to study the genetic differences in plants and animals. DNA fingerprinting, gene
mapping and polymorphic studies have been tremendously benefited from PCR. One
variation of PCR is the random amplified polymorphic DNA (RAPD), which generates
DNA fingerprinting with a single synthetic oligo-nucleotide primer (Williams et al., 1990).
RAPDs are inherited in a simple Mendelian fashion and are usually dominant markers.
Gene mapping using RAPD markers has several advantages over RFLPs. RAPD procedure
is less expensive, faster, requires less amount of DNA (0.5 to 50 ng) and does not involve
the radioisotopes. The objective of the present study was to find out genetic diversity and
genetic relationship among six varieties of chickpea and their F1 crosses.
The DNA fingerprinting will provide an additional proof for the varieties under
study. This procedure of DNA fingerprinting comprised the following steps.
28
(b) Plant sample
Seeds of six parental genotypes i.e., CM-98, AUG-786, Bittal-98, Balksar-2000,
Wanhar-2000 and Punjab-2000 and their F1 single crosses were grown in earthen pots and
supplied with optimum amount of water and nutrition through Hoagland solution. Later
leaf tissues were collected for DNA extraction.
(c) DNA Extraction
The total genomic DNA was extracted by CTAB method (Doyle and Doyle 1990).
Turned on the water bath and settled at 650C and preheated 2xCTAB. Autoclaved pestle
and mortar. Cut 4-5 young leaves, washed with distilled water, dried and grinded into a
very fine powder in liquid nitrogen. Transferred the paste to a 15 ml blue cap tube (Falcons
tube). Added 15 ml of hot (650C) CTAB (with 1 per mercap to ethanol) to the tube before
frozen powder starts thawing. Mixed gently by inverting a tube for several times and
incubated at 650C for half an hour. Added 15 ml of chloroform isoamyl alcohol (24: 1) and
mixed gently by inverting a tube to form an emulsion. Centrifuge material for 10 minutes at
9000 rpm. The supernant transferred to a new falcon tube. Repeated the above two steps.
Added 0.6 volumes or 60 % Isopropanol (pre-chilled) and mixed gently to precipitate the
DNA. Centrifuged at 9000 rpm for 5 minutes and discarded the supernatant. Took the
pellet and washed with 70 % ethanol twice or thrice. Air dried the pellet and resuspended
in 0.5 ml 0.1x TE buffer or d3H2O. The suspension transferred into eppendrof tube from
blue cap tube. Then added 7ul of RNAase and incubated for 1 hour at 37 C0, mixed gently
before incubation. Equal volume of chloroform isoamyle alcohol (24:1) was added and
mixed gently. Added 10ul of 3 M NaCl and mixed gently. Precipitate the DNA with
absolute ethanol (cool, 2 volume normally 7000 ul). Span at 13000 rpm for 10 minutes,
discarded the supernant, washed the pellet with 70 % ethanol, air dried and resuspended in
0.1xTE buffer (10 mM Tris.Cl, 0.1 mM EDTA, pH 8.0) or d3H2O and measure the
concentration of DNA at 260 nm.
(d) Estimation of DNA concentration
For the concentration of total genomic DNA of 21 samples, absorbance was
measured at 260 nm on spectrophotometer. A solution with an O.D.260 contains 50µg of
DNA per millimeter where, DNA quantity is calculated by the following formula.
29
DNA Con. in µg / ml = Absorbance at 260 nm x Dilution factor x 50
(e) RAPD analyses
DNA concentration in the working solution of approximately 12ngμl-1
in d3H2O was
confirmed by spectrophotometer. For RAPD analysis (Williams et al. 1990), concentration
of genomic DNA, 10xPCR buffer with (NH4)2SO4, MgCl2, dNTPs (dATP, dCTP, dGTP,
dTTP), 10-mer random primers and Taq DNA polymerase were optimized. The 10 base
oligo-nucleotide primers obtained from Gene Link Company (USA) were used for the
amplification of the genomic DNA. While Taq polymerase together with buffer, MgCl2,
dNTPs, gelatin, were purchased from Fermentas. DNA amplification reactions were
performed in a thermal cycler (Eppendorf AG No. 5333 00839). The PCR profile was
followed as, one cycle of 94 ºC for 5 minutes, 40 cycles of, 94ºC for 1 minute, 36ºC for 1
minute 72 ºC for 2 minutes and final extension for 10 minutes at 72 ºC.
(f) Scoring and analysis of RAPD data
Amplification products were analyzed by electrophoresis in 1.2 % (w/v) agarose
gels and were detected by staining the gel with ethidium bromide (10 ng/100ml of agarose
solution in TBE). All visible and unambiguously score-able fragments amplified by
primers were scored under the heading of total score-able fragments. Amplification profiles
of all the 21 chickpea genotypes were compared with each other and bands of DNA
fragments were scored as present ‘1’ or absent ‘0’. The data of the primers were used to
estimate the similarity on the basis of the number of shared amplification products (Nei and
Li, 1979). Similarity coefficients were utilized to generate a dendrogram by means of un-
weighted pair group method of arithmetic means (UPGMA).
4. Chickpea wilt inheritance
The chickpea wilt was first mentioned in this region in 1918 (Nene, 1980).
Fusarium wilt is reported, outside, Pakistan, from India, Burma, Russia, Ethopia, Malawi,
Tunisia, Spain, U.S.A., Mexico and Peru (Allen, 1983). To study inheritance of resistance
to chickpea wilt, six resistant (class 0) parents viz., CM-98, AUG-786, Bittal-98, Balksar-
2000, Wanhar-2000, Punjab-2000 and one disease susceptible parent viz., AUG-424 were
crossed during February 2006. To ascertain their reaction classes and genetic difference,
AUG-424 was also crossed with each other. But the cross was not successful because of
30
severe fungal effect of pathogen. The six successful crosses were manipulated and F1, F2
progenies were raised and evaluated for disease reaction under wilt sick bed during the
seasons of 2006-07 to 2008-09, respectively. To ensure uniform incidence of the disease
under field conditions, artificial sickbed was prepared. The F3 progenies were obtained
from random F2 plants grown under wilt free conditions.
Chi-square Test
In all genetic problems, genetic behavior is observed by phenotype of an organism
in a given environment. Experimental results seldom confirm exactly to the expected
phenotypic ratio due to chance events in the meiotic production of gametes, random union
of these gametes in fertilization and sample size taken from a population. Chi-square test is
used successfully to convert deviations from expected values into the probability of such
inequalities occurring by chance (Fisher, 1948; Gomez and Gomez, 1984).
31
Chapter-4
RESULTS AND DISCUSSION
Analysis of variance
Mean squares from the analysis of variance for the traits under study of Cicer
arietinum L. genotypes are presented in Table.2 from six parents and a set of complete
diallel cross including F1s and reciprocals. Analysis of variance indicated highly significant
differences among the genotypes for all the traits under study.
Presence of significant genotype differences for all the traits allowed to proceed for
further analysis of variance as prescribed by Steel and Torrie (1994). The biometrical
analysis comprised diallel analysis (Hayman, 1954a, b and Mather and Jinks, 1982),
combining ability analysis (Griffing, 1956) and chi-square test (William, 1969, Burns,
1983, Gomez and Gomez, 1984).
1. Diallel analysis
Diallel analysis assumptions and tests for their validity
Diallel analysis assumptions include regular diploid behavior at meiosis, no
significant difference between reciprocal crosses, homozygous parents, absence of epistasis
and multiple allelism and independent gene distribution. Highly significant differences
among the genotypes (Table.2) suggested that detailed analysis of gene action and
combining ability were necessary. To fulfill the assumptions of absence of epistasis, no
multiple allelism and independent gene distribution, data were subjected to two tests viz.,
analysis of variance to test the constancy of Wr-Vr over arrays and a joint regression
analysis of Wr on Vr, were carried out. The significant mean squares for analysis of Wr +
Vr and Wr-Vr suggested partial failure of the assumptions for most of the traits. However
the joint regression analysis fulfilled the requirements of additive-dominance model for
yield and its components except harvest index (Table.3). The regression coefficient for
harvest index was non-significant indicating inadequate assumption in this experiment.
Therefore no further analysis was computed on this trait. Such partial failure of
assumptions is of little value to interpret diallel statistics either in term of distribution of
32
Table.2. Analysis of variance for yield and its various components.
Character
MEAN SQUARE
Genotypes Replications Error
d.f = 35 d.f = 2 d.f = 70
1 Days taken to flowering 25.876** 1.398NS
0.598
2 Plant height 76.904** 0.342NS
0.608
3 Number of primary branches
per plant
0.115** 0.065NS
0.001
4 No. of secondary branches per
plant
0.514** 0.001NS
0.003
5 Days taken to maturity 69.996** 0.003NS
0.695
6 Total weight of plant 82.451** 0.287NS
0.183
7 Number of pods per plant 84.513** 0.0001NS
2.077
8 Number of seeds per pod 0.005** 0.607* 0.004
9 100-seed weight (g) 2.836** 0.019NS
0.010
10 Grain yield per plant 22.742** 0.046NS
4.444
11 Harvest index % 19.684** 0.595NS
0.858
** = Significant at 1% probability level NS = Non-significant
33
Table 3. Adequacy test of additive dominance model for 6×6 diallel
cross of Cicer arietinum L.
Characters Regression
Analysis Analysis of Array Variance
Remarks
b= 0 b=1 Wr+Vr Wr-Vr 1 Number of days
taken to flowering
3.049* 1.391NS 18.732** 2.252NS Adequate
2 Number of primary branches per plant
13.412*
* -1.043NS 134.032** 8.025** Partial
adequate
3 No. of secondary branches per plant
2.844** 0.131NS 275.566** 141.912**
Partial adequate
4 Plant height (cm) 6.719** -1.093NS 20.128** 1.303NS Adequate
5 Number of days taken to maturity
2.841* 0.103NS 31.147** 10.014*
* Partial adequate
6 Total weight of plant (g)
3.080* 0.923NS 234.640** 313.994**
Partial adequate
7 Number of pods per plant
3.222* 0.598NS 19.368** 8.858** Partial adequate
8 Number of seeds per pod
2.793* 0.457NS 13.099** 1.121NS Adequate
9 100-seed weight (g)
2.818* 0.571NS 367.421** 483.833**
Partial adequate
10 Grain yield per plant (g)
3.044* 0.633 NS 36.243** 14.322*
* Partial adequate
11 Harvest index % 0.951NS 2.589NS 23.519** 20.374*
* Inadequate
* = Significant at 5% probability level ** = Significant at 1% probability level NS = Non-significant
34
alleles, additive and dominance genetic variance and number of genes. Partial failure of the
assumptions revealed a more complex genetic system than that described by the theoretical
model and it is possible to make estimates of genetical components for such complex
system (Hayman, 1954a and Dickinson and Jinks, 1956; Johnson and Askel, 1964; Baker,
1978 and Wilson et al. 1978).
Number of days taken to flowering
Diallel analysis for days taken to flowering (Table.4) revealed that both additive (a)
and dominance (b) components were highly significant (P≤ 0.01) suggesting the
importance of both additive and dominance variations in the inheritance of this trait.
However, the component b1 was non-significant indicating that the directional dominance
was absent among parents. The highly significant mean squares for b2 and b3 revealed the
symmetry of gene distribution among parents and importance of specific gene interaction
for days taken to flowering, respectively. Maternal effects (c) were non-significant (P≤
0.01). Reciprocal effects (d) were significant. The items b, b1, b2 and b3 after retesting
against d item were highly significant (P≤ 0.01) indicating the importance of reciprocal
effects in the inheritance of the trait.
The genetic components of variation for days taken to flowering (Table.5) revealed
significant additive (16.256) and dominance (14.163) effects. D was greater than H1
indicating that additive effects were more important for the inheritance of this trait. The
closeness of H1 and H2 values indicated that positive and negative alleles were equal
among the parents. H2/4H1 ratio (0.225) also showed similar distribution of positive and
negative genes. The value of F component of variation was positive and non-significant
which indicated that dominant and recessive alleles were statistically at par. The ratio of
dominant to recessive genes (1.341) indicated that dominant genes were more frequent than
the recessive genes in the parents. The non-significant values of ‘h2’ and environmental
effect (E) indicated that net dominant effects and environmental effects had little value for
the expression of this trait. Average degree of dominance was less than 1 (0.933) showing
partial dominance type of gene action for trait expression and this was verified by the slope
35
Table 4. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for number of days taken to flowering in chickpea.
ITEMS d.f MS F Ratio Re-testing against c & d
C d
a 5 119.841 475.838** - -
b 15 19.742 23.992** - 25.382**
b1 1 17.424 11.351 - 22.402**
b2 5 6.993 8.582** - 8.990**
b3 9 27.081 36.200** - 34.819**
c 5 0.511 0.541
d 10 0.778 2.980*
Blocks 2 1.398
B × a 10 0.252
B × b 30 0.823
B × b1 2 1.535
B × b2 10 0.815
B × b3 18 0.748
B × c 10 0.944
B × d 20 0.261
Bl.Int.
Total
70
107
0.598
* = Significant at 5% level of probability ** = Significant at 1% level of probability
36
Table 5. Estimates of genetic components of variation for number of days taken
to flowering in chickpea.
COMPONENTS ESTIMATES
D 16.256* ±1.267
H1 14.163* ±3.215
H2 12.747* ±2.872
F 4.425NS ±3.094
h 3.112NS ±1.933
E 0.207NS ±0.483
√H1/D 0.933
H2/4H1 0.225
√4DH1+ F/ √4DH1- F 1.341
h2 ns 0.6612
h2 bs 0.9793
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
37
Fig.1. Wr/Vr graph for days taken to flowering.
38
of the regression line, which intercepted the Wr-axis above the origin (Fig.1). These
findings are in agreement with the observations of Sandhu and Mandal (1989), Singh et al.
(1992), Durga et al. (2007), Bicer and Sakar (2008) and Biranvand et al. (2008) who
reported additive and dominance type of gene action for this trait. Girase and Deshmukh
(2000) observed duplicate type of gene action in the expression days to flowering. Non-
additive type of gene action was reported by Patil et al. (2006) and Meena et al. (2006).
High heritability with low genetic advance was reported by Arshad et al. (2004). Derya et
al. (2006) reported 5.47% broad sense heritability and 66 % narrow sense heritability for
this character, indicated that more than 66 % of the genetic variation was of additive type.
Graphical analysis (Fig.1) for the character depicted that genotype Bittal-98
possessed the maximum number of dominant genes for days taken to flower, on the other
hand genotype CM-98 and Balksar-2000 carried maximum number of recessive genes. The
remaining parents falling in between were intermediate for distribution of the genes.
Number of primary branches per plant
Diallel analysis for number of primary branches per plant (Table.6) revealed highly
significant (P≤ 0.01) mean squares (0.69) due to additive (a) gene effects. The overall
dominance component b was much smaller (0.03) but highly significant showing the
importance of dominant (b) effects. The item b1 and b2 were highly significant (P≤ 0.01)
indicating the presence of unidirectional dominance of the genes and asymmetry of gene
distribution among parents respectively. The item b3 was highly significant which means
that specific gene interaction was present. Maternal effects (c) were non-significant at (P≤
0.05) indicating the absence of maternal effect. Reciprocal effects (d) were non-significant
indicating the absence of reciprocal effects. The presence of differences among reciprocal
crosses item b retested against d mean square.
The estimation of genetic components of variation for number of primary branches
per plant (Table.7) revealed that both additive (0.079) and dominant component (0.027) of
variations were significant. However ‘D’ component was much greater than H1 indicating
39
Table 6. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for number of primary branches per plant in chickpea.
ITEMS d.f MS F Ratio a 5 0.689 973.734
**
b 15 0.035 104.263**
b
1 1 0.115 2328.731
**
b2 5 0.019 81.272
**
b3 9 0.035 82.658
**
c 5 0.001 0.889 d 10 0.005 4.791 Blocks 2 0.001 B ×a 10 0.001 B × b 30 0.001 B × b
1 2 0.001
B × b2 10 0.001
B × b3 18 0.001
B × c 10 0.001 B × d 20 0.001 Bl.Int. 70 0.001 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
40
Table 7. Estimates of genetic components of variation for number of primary
branches per plant in chickpea.
COMPONENTS ESTIMATES
D 0.079* ±0.001
H1 0.027
* ±0.004
H2 0.023
* ±0.003
F 0.007NS
±0.004
h2 0.021
* ±0.002
E 0.0002NS
±0.0006
√H1/D 0.586
H2/4H
1 0.212
√4DH1+ F/ 4DH
1- F 1.162
h2 ns 0.8647
h2
bs 0.9957
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
41
Fig.2. Vr/Wr graph for number of primary branches per plant.
42
additive genes controlling for this trait. The estimates of H1 and H2 and H2/4H1 ratio 0.212
suggested the equal distribution of positive and negative alleles among the parents. The
value of F component of variation (0.007) was positive and non-significant which indicated
that dominant and recessive alleles were statistically at par. Ratio of dominant to recessive
genes (1.162) was slightly greater than 1 showing more number of dominant than recessive
alleles.
The dominance effect of heterozygous loci (0.021) for this trait was significant. The
values of average degree of dominance (0.586) was less than 1, that was above the graphic
origin (Fig.2) intercept the regression line suggested that additive gene action with partial
dominance for this trait. Environmental effects (E) was non-significant, indicated that
environmental effects have little value for the expression of the trait. The estimates of
narrow and broad sense heritability were high. Narrow sense heritability was (86.5%) of
broad sense heritability and additive type. Additive and dominant gene effects were
reported by Singh et al. (1992) and Bicer and Sakar (2008). Non-additive type of gene
action was reported by Meena et al. (2006).
Graphical analysis (Fig.2) for the character depicted that genotype CM-98 possessed
maximum number of dominant genes for number of primary branches per plant, on the
other hand the genotype AUG-786 carried maximum number of recessive genes. The
remaining parents falling in between two were intermediate for distribution of the genes.
Number of secondary branches per plant
Diallel analysis for number of secondary branches per plant (Table.8) revealed that
both additive (a) and dominance (b) components were highly significant (P≤ 0.01)
suggesting the importance of both additive and dominance effects for the inheritance of
number of secondary branches per plant. However, the component b1 was non-significant
indicating that the directional dominance was absent among parents. The highly significant
mean squares for b2 and b3 revealed the symmetry of gene distribution among parents and
importance of specific gene interaction for number of secondary branches per plant
respectively. Maternal effects (c) and reciprocal effects (d) were non-significant (P≤ 0.01).
43
Table 8. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for number of secondary branches per plant in chickpea.
ITEMS d.f MS F Ratio a 5 2.529 956.931
**
b 15 0.354 87.233**
b
1 1 0.109 10.383
b2 5 0.164 37.641
**
b3 9 0.487 153.473
**
c 5 0.003 1.762 d 10 0.003 1.172 Blocks 2 0.003 B × a 10 0.003 B × b 30 0.004 B × b
1 2 0.010
B × b2 10 0.004
B × b3 18 0.003
B × c 10 0.002 B × d 20 0.002 Bl.Int. 70 0.003 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
44
Table 9. Estimates of genetic components of variation for number of secondary
branches in chickpea.
COMPONENTS ESTIMATES
D 0.459* ±0.044
H1 0.270
* ±0.113
H2 0.234
* ±0.101
F 0.214* ±0.108
h2 0.020
* ±0.068
E 0.001NS
±0.017
√H1/D 0.767
H2/4H
1 0.217
√4DH1+ F/ 4DH
1- F 1.874
h2 ns 0.7021
h2
bs 0.9949
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
45
Fig.3. Vr/Wr graph for number of secondary branches per plant.
46
The genetic components of variation for number of secondary branches per plant
(Table.9) revealed significant additive (0.459) and dominance (0.270) variations. D was
greater than H1 indicating that dominance effects were more important for this trait. The
closeness of H1 and H2 values indicated that positive and negative alleles were equal
among the parents. H2/4H1 ratio (0.217) also showed similar distribution of positive and
negative genes. The value of F component of variation was positive and significant
indicated that dominant and recessive alleles were statistically at par. The significant values
of ‘h2’ indicated the importance of net dominant effects. Non-significant environmental
effect (E) indicated that environmental effects have little value for the expression of the
trait. Average degree of dominance was less than 1 (0.767) showing partial dominance type
of gene action for trait expression and this was verified by the slope of the regression line,
which intercepted the Wr-axis above the origin (Fig.3).
Narrow sense heritability for the character indicated that more than 70 % of the
genetic variation was of additive type. Additive, dominant gene effects were reported by
Singh et al. (1992) and Bicer and Sakar (2008) and Sreelakshmi et al. (2010). Additive type
of gene action was reported by Patil et al. (2006) and Arshad et al. (2004). Non-additive
type of gene action was reported by Meena et al. (2006). Girase and Deshmukh (2000)
observed duplicate type of gene action in the expression number of secondary branches per
plant.
Graphical analysis (Fig.3) for the character depicted that genotype Balksar-2000
possessed maximum number of dominant genes for number of secondary branches per
plant, on the other hand genotype Punjab-2000 carried maximum number of recessive
genes. The remaining parents falling in between were intermediate for distribution of the
genes.
Plant Height (cm)
Diallel analysis for plant height (Table.10) revealed highly significant (P≤ 0.01)
mean squares (399.7) due to additive (a) gene effects. The overall dominance component b
was much smaller (24.1) but highly significant showing the importance of dominant (b).
The item b1 was significant (P≤ 0.05) indicating the presence of unidirectional dominance
of the genes. The item b2 was highly significant showing the asymmetry of gene
distribution among parents. The item b3 was non-significant which means that specific gene
47
interaction was absent. Maternal effects (c) and reciprocal effects (d) were significant (P≤
0.01) indicating the presence of maternal as well as reciprocal effects. The presence of
significance differences among maternal and reciprocal crosses item ‘a’ was retested
against c and b item against d mean squares. After retesting the ‘a’ item against ‘c’
remained highly significant indicating that maternal effects had not influence additive
genetic effects. After retesting d items, b and its subdivisions b1, b2 and b3 changed into
non-significant showing the influence of reciprocal effects on dominance effects.
The estimation of genetic components of variation for plant height (Table.11)
revealed that both additive (43.5) and dominant component (17.1) of variation were
significant. However D component was much greater than H1 indicated additive genes
controlling for this trait. The estimates of H1, H2 and H2/4H1 ratio 0.229 suggested the
equal distribution of positive and negative alleles among the parents. The value of F
component of variation was positive and non-significant which indicated that dominant and
recessive alleles were statistically at par. Ratio of dominant to recessive genes (1.020) was
slightly greater than 1 showing equal number of dominant and recessive alleles.
The dominance effect of heterozygous loci (4.67) for this trait was non-significant.
The value of average degree of dominance (0.626) was less than 1, suggested that additive
gene action with partial dominance, and the graphical analysis (Fig.4) also displayed
similar trend of gene action for trait expression. Environmental effects (E) were non-
significant, indicated that no influence of environment for the expression of the trait. The
estimates of narrow and broad sense heritability were high. Narrow sense heritability was
(84.4%) of broad sense heritability. Significant additive and dominance gene action was
also reported by Singh et al. (1992) and Beyran et al. (2008). Significant additive gene
action were reported by Singh et al. (1991, Patil et al. (2006), Bicer and Sakar (2008).
Non-additive type of gene action was reported by Meena et al. (2006). Biranvand et al.
(2008) and Beyran et al. (2008) estimated genetic parameters D, H1 and H2, indicated that
additive variance component was greater than non-additive variance components, narrow
sense heritability was medium and Wr/Vr graphs reflected partial dominance for plant
height.
48
Table 10. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for Plant height (cm) in chickpea.
ITEMS d.f MS F Ratio Re-testing against c & d
c d a 5 399.674 512.646
** 14.029
** -
b 15 24.050 36.550**
- 1.265 b
1 1 25.785 37.330
* - 1.357
b2 5 6.998 9.531
** - 0.368
b3 9 33.331 54.460 - 1.754
c 5 28.489 38.556**
d 10 19.005 49.942
**
Blocks 2 0.065 B × a 10 0.780 B × b 30 0.658 B × b
1 2 0.691
B × b2 10 0.734
B × b3 18 0.612
B × c 10 0.739 B × d 20 0.380 Bl.Int. 70 0.608 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
49
Table 11. Estimates of genetic components of variation for plant height (cm) in
chickpea.
COMPONENTS ESTIMATES
D 43.465* ±1.889
H1 17.062
* ±4.796
H2 15.639
* ±4.284
F 0.546NS
±3.615
h2 4.665
NS ±2.884
E 0.197NS
±0.721
√H1/D 0.626
H2/4H
1 0.229
√4DH1+ F/ 4DH
1- F 1.020
h2 ns 0.8437
h2
bs 0.9925
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
50
Fig.4. Vr/Wr graph for plant height.
51
Wr/Vr graphs reflected partial dominance for plant height. Graphical analysis
(Fig.4) depicted that genotype Bittal-98 possessed maximum number of dominant genes for
plant height, on the other hand genotypes CM-98, AUG-786, Wanhar-2000, Punjab-2000
and Balksar carried the maximum number of recessive genes.
Number of days taken to maturity
Analysis of variance for number of days taken to maturity (Table.12) revealed that
both additive (a) and dominant (b) component were highly significant (P≤ 0.01). However,
the component b1 were non-significant showing the absence of directional dominance
deviations of the genes. Significant b2 item indicated symmetry of gene distribution among
the parents. The item b3 was non-significant displaying unimportance of specific gene
interaction. Maternal effects (c) were non-significant and had no importance in trait
expression. Reciprocal effects (d) were significant indicating the importance of reciprocal
effects. After retesting of d against b, b1, b2 and b3 it shown the same trend and become
highly significant.
The estimates of genetic components of variation for days taken to maturity (Table.13)
revealed significant additive (37.43) and dominance (42.82) variation. D was smaller than
H1 indicating that dominance variations were more important for this trait. The closeness of
H1 and H2 values suggested that positive and negative alleles were equal among the
parents. H2/4H1 ratio (0.218) also showed similar distribution of positive and negative
genes.
The value of F component of variation was positive and non-significant indicated
that dominant and recessive alleles were statistically at par. The ratio of dominant to
recessive genes (1.222) also showed that dominant and recessive genes were equally
controlled the character. The values of ‘h2’ and environmental effect (E) were non-
significant, indicated that no influence of net dominant effects due to heterozygous loci and
environmental effects for the expression of the trait. Average degree of dominance was
greater than1 (1.069) shown over dominance type of gene action and the graphical analysis
(Fig.5) also displayed similar trend of gene action for trait expression. Additive, dominant
gene effects were reported by Singh et. al. (1992), Bicer and Sakar (2008) and Biranvand
(2008).
52
Table 12. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for number of days taking to maturity in chickpea.
ITEMS d.f MS F Ratio Re-testing against c & dc d
a 5 315.107 358.277**
- b 15 56.676 81.552
** 38.938
**
b1 1 33.252 9.260 22.845
**
b2 5 25.392 54.958
** 17.445
**
b3 9 76.659 128.759 52.667
**
c 5 1.922 2.601 d 10 1.455 2.927* Blocks 2 0.342 B × a 10 0.879 B × b 30 0.751 B × b
1 2 3.591
B × b2 10 0.462
B × b3 18 0.595
B × c 10 0.739 B × d 20 0.497 Bl.Int. 70 0.695 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
53
Table 13. Estimates of genetic components of variation for number of days taken
to maturity in chickpea.
COMPONENTS ESTIMATES
D 37.434* ±4.057
H1 42.818
* ±10.298
H2 37.327
* ±9.200
F 7.989NS
±9.911
h2 6.031
NS ±6.192
E 0.228NS
±1.548
√H1/D 1.069
H2/4H
1 0.218
√4DH1+ F/ 4DH
1- F 1.222
h2 ns 0.5762
h2
bs o.9915
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
54
Fig.5. Wr/Vr graph for number of days taken to maturity.
55
Non-additive type of gene action was reported by Meena et al. (2006). High
heritability with low genetic advance was reported by Arshad et al. (2004). Girase and
Deshmukh (2000) observed duplicate type of gene action in the expression days to
maturity. The estimate of narrow sense heritability for the character indicated (58%) the
genetic variation was of dominance nature.
Graphical analysis (Fig.5) for the character depicted that genotype AUG-786
possessed maximum number of dominant genes for days taken to maturity, on the other
hand genotype Punjab-2000 carried maximum number of recessive genes. The remaining
parents falling in between two were intermediate for distribution of the genes.
Total weight of plant (g)
Diallel analysis for total weight of plant (Table.14) revealed highly significant (P≤
0.01) mean squares due to additive (a) and dominant gene effects. The overall dominance
component b was much smaller (27.59) indicated the pre-dominance of additive effects.
The item b1 and b2 were highly significant (P≤ 0.01) indicating the presence of
unidirectional dominance of the genes and the asymmetry of gene distribution among
parents respectively. The item b3 was highly significant which indicated that specific gene
interaction was present. Maternal effects (c) were not detected, whereas reciprocal effects
(d) were highly significant indicating the importance of reciprocal crosses. The presence of
differences among reciprocal crosses item d were retested against b mean squares. After
retesting non-additive item b and its subdivisions b2 and b3 remained significant showing
the influence of reciprocal effects on dominance effects.
The estimation of genetic components of variation for total weight of plant
(Table.15) revealed that both additive (56.72) and dominant component (23.07) of variation
were significant. However D component was much greater than H1 indicating additive gene
controlling was predominating for this trait. The estimates of H1 and H2 and H2/4H1 ratio
0.198 suggested the unequal distribution of positive and negative alleles among the parents.
56
Table 14. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for total weight of plant (g) in chickpea.
ITEMS d.f MS F Ratio Re-testing against c & d
c d a 5 491.394 2570.254
** - -
b 15 27.590 220.510**
- 21.457**
b1 1 0.832 149.315
** - 0.647
b2 5 21.844 187.981
** - 16.989
**
b3 9 33.754 235.466
** - 26.252
**
c 5 0.422 2.582 d 10 1.286 4.648
**
Blocks 2 0.607 B × a 10 0.191 B × b 30 0.125 B × b
1 2 0.005
B × b2 10 0.116
B × b3 18 0.143
B × c 10 0.163 B × d 20 0.277 Bl.Int. 70 0.183 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
57
Table 15. Estimates of genetic components of variation for total weight of plant in
chickpea.
COMPONENTS ESTIMATES
D 56.724* ±3.968
H1 23.073
* ±5.072
H2 18.263
* ±4.998
F 6.958NS
±5.693
h2 0.118
NS ±3.056
E 0.065NS
±1.514
√H1/D 0.638
H2/4H
1 0.198
√4DH1+ F/ 4DH
1- F 1.213
h2 ns 0.8549
h2
bs 0.9980
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
58
Fig.6. Vr/Wr graph for total weight of plant.
59
The value of F component of variation (6.958) was positive and non-significant which
indicated that dominant and recessive alleles were statistically at par. Ratio of dominant to
recessive genes was greater than 1 (1.213) showed the greater frequency of dominant gene.
Net dominance effects due to heterozygous loci (0.118) were significant for this trait. The
values of average degree of dominance (0.638) were less than 1, that was above the graphic
origin intercept the regression line (Fig.6) suggested that additive gene action with partial
dominance.
Environmental effects (E) were non-significant, indicated unimportance of
environment for the expression of the trait. The estimates of narrow and broad sense
heritability were high. Narrow sense heritability was (85.5%) of broad sense heritability
and additive in nature. Additive gene effects were important reported by Singh and Bains
(1985) Tewari and Pandey (1986) and Arshad et al. (2004). Venkatraman (2007) indicated
the predominant role of additive gene action on biomass. Non-additive type of gene action
was reported by Meena et al. (2006). Singh and Bains (1982) observed both additive and
non-additive gene action were important for total weight of plant. Hussain et al. (1990)
found mode of inheritance appeared to be over-dominant for total plant weight.
Graphical analysis (Fig.6) for total plant weight depicted that genotype Balksar-
2000 possessed maximum number of dominant genes, on the other hand genotype CM-98
and AUG-786 carried the maximum number of recessive genes. The remaining parents
falling in between two were intermediate for distribution of the genes.
Number of pods per plant
A perusal of results for number of pods per plant (Table.16) revealed highly
significant (P≤ 0.01) mean squares due to additive (a) and dominant (b) genetic effects.
The overall dominance component b was smaller (77.54) indicated the pre-dominant role of
additive genetic effects. The item b1 and b2 were highly significant (P≤ 0.01) indicating the
presence of unidirectional dominance of the genes and the asymmetry of gene distribution
among parents respectively. The item b3 was highly significant which means that specific
gene interaction was present. Maternal effects (c) and reciprocal effects (d) were non-
significant at (P≤ 0.05) indicating that retesting was not required.
60
The estimation of genetic components of variation for number of pods per plant
(Table.17) revealed that both additive (42.94) and dominant component (57.07) of variation
were significant. However ‘D’ component was smaller than H1 indicating predominant
genes controlling this trait. The estimates of H1 and H2 and H2/4H1 ratio 0.220 suggested
the equal distribution of positive and negative alleles among the parents. The value of F
component of variation (10.35) was positive and non-significant which indicated that
dominant and recessive alleles were statistically at par. Ratio of dominant to recessive
genes (1.23) was slightly greater than 1 showing more dominant genes than recessive
genes. The net dominance effect due to heterozygous loci (0.021) was also significant. The
value of average degree of dominance (1.153) was greater than 1, that was slightly below
the graphic origin (Fig.7) intercept the regression line suggested over dominance types of
gene action for this trait.
Environmental effects (E) was non-significant, indicated that environmental effects
have little value for the expression of the trait. The estimates of narrow and broad sense
heritability were high. Narrow sense heritability was (59.7%) of broad sense heritability
and dominance in nature. Additive and dominance gene action was reported by Singh et al.
(1992), Chaturvedi et al. (1997) and Beyran et al. (2008). Significant additive gene action
was reported by Singh et al. (1982), Tewari and Pandey (1986), Yadav et al. (1987), Durga
et al. (2007), Bicer and Sakar (2008) and Sreelakshmi et al. (2010). Non-additive type of
gene action was reported by Kulkarni (2001), Sarode et al. (2001), Patil et al. (2006) and
Meena et al. (2006). Biranvand et al. (2008) estimated genetic parameters D, H1 and H2,
indicated that additive variance component was less than non-additive variance
components, narrow sense heritability was low and Wr/Vr graphs reflected over dominance
for number of pods per plant.
Graphical analysis (Fig.7) for the character depicted over-dominant types of gene
action. The genotype Balksar-2000 and Bittal-98 possessed maximum number of dominant
genes for number of pods per plant, on the other hand genotype Punjab-2000 carried
maximum number of recessive genes.
61
Table 16. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for number of pods per plant in chickpea.
ITEMS d.f MS F Ratio a 5 355.941 163.721
**
b 15 77.539 32.151**
b
1 1 113.896 140.100
**
b2 5 32.309 8.408
**
b3 9 98.628 54.963
**
c 5 0.689 0.305 d 10 1.172 0.815 Blocks 2 0.287 B × a 10 2.174 B × b 30 2.412 B × b
1 2 0.813
B × b2 10 3.842
B × b3 18 1.794
B × c 10 2.255 B × d 20 1.439 Bl.Int. 70 2.077 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
62
Table 17. Estimates of genetic components of variation for number of pods per
plant in chickpea.
COMPONENTS ESTIMATES
D 42.942* ±5.669
H1 57.070
* ±3.391
H2 50.341
* ±4.856
F 10.348NS
±3.849
h2 20.716
* ±2.653
E 0.676NS
±2.164
√H1/D 1.053
H2/4H
1 0.220
√4DH1+ F/ 4DH
1- F 1.233
h2 ns 0.5972
h2
bs 0.9795
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
63
Fig.7. Vr/Wr graph for number of pods per plant.
64
The remaining parents fall near to Punjab-2000, which has recessive distribution of the
genes.
Number of seeds per pod
Analysis of variance for number of seeds per pod presented in Table.18 revealed
that both additive (a =0.013) dominant (b=0.007) component were highly significant (P≤
0.01) suggested its importance in trait expression. The overall dominance component b was
smaller than a indicated the pre-dominant role of additive genetic effects. However, the
component b1 (0.001) was non-significant showing the absence of directional dominance
deviations of the genes. The item b2 was significant indicating symmetry of gene
distribution among parents. The item b3 was significant displaying importance of specific
gene interaction. Maternal effects (c) was non-significant have no importance and
reciprocal effects (d) was significant have importance in trait expression. The presence of
differences among reciprocal crosses item d were retested against b items mean squares.
After retesting non-additive item b and its subdivision b3 remained significant showing the
influence of reciprocal effects on dominance effects.
The estimates of genetic components of variation for number of seeds per pod
(Table.19) revealed significant additive (0.0014) and dominance (0.0046) effects. D was
smaller than H1 indicating that dominance effects were pre-dominating for this trait. The
closeness of H1 and H2 values suggested that positive and negative alleles were equal
among the parents. H2/4H1 ratio (0.228) also showed similar distribution of positive and
negative genes.
The value of F component of variation was positive and non-significant which
indicated that dominant and recessive alleles were statistically at par. Ratio of dominant to
recessive genes (1.163) also showed that dominant genes were more frequent and
controlled the character. The values of ‘h2’ (0.0002) and ‘E’ (0.0001) were non-significant,
indicated that net dominant effects due to heterozygous loci and environmental effects have
little value for the expression of the trait. Average degree of dominance was greater than 1
(1.813) showing over dominance type of gene action for trait expression.
65
Table 18. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for number of seeds per pod in chickpea.
ITEMS d.f MS F Ratio Re-testing against
c & d c c d
a 5 0.013 24.092**
-
b 15 0.007 18.546**
- 7.277**
b1 1 0.001 1.159 - 1.353
b2 5 0.002 10.687
** - 2.415
b3 9 0.010 26.682
** - 10.637**
c 5 0.001 2.697
d 10 0.001 3.058*
Blocks 2 0.0001 B × a 10 0.0005 B × b 30 0.0004 B × b
1 2 0.0011
B × b2 10 0.0002
B × b3 18 0.0004
B × c 10 0.0004 B × d 20 0.0003 Bl.Int. 70 0.0004 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
66
Table 19. Estimates of genetic components of variation for number of seeds per
pod in chickpea.
COMPONENTS ESTIMATES
D 0.0014* ±0.0004
H1 0.0046
* ±0.0010
H2 0.0042
* ±0.0009
F 0.0004NS
±0.0010
h2 0.0002
NS ±0.0006
E 0.0001NS
±0.0001
√H1/D 1.8134
H2/4H
1 0.2278
√4DH1+ F/ 4DH
1- F 1.1627
h2 ns 0.3785
h2
bs 0.9348
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
67
Fig.8. Vr/Wr graph for number of seeds per pod.
68
The estimate of narrow sense heritability for the character indicated (38%) the
genetic variation was of dominance nature. Additive and dominance gene action was
reported by Chaturvedi et al. (1997), Sarode et al. (2001) and Biranvand (2008). Additive
gene action was reported by Yadav et al. (1987), Singh et al. (1992) and Beyran et al.
(2008). Both additive and non-additive gene actions important with additive predominating
were observes by Singh and Bains (1982) and Malhotra et al. (1983). Non-additive type of
gene action was reported by Meena et al. (2006). Martinez et al. (1978) observed full
dominance in a positive sense in seeds per pod. Patil et al. (1998) observed predominance
of epistatic gene action. Sandhu and Mandal (1989) and Annigeri et al. (1996) revealed the
predominance of additive variance for seeds per pod. Kumar et al. (2001) revealed the
predominance of non-additive variance for this trait.
Graphical analysis (Fig.8) for the character depicted over dominant types of gene
action. The genotype AUG-786, Balksar-2000 and Punjab-2000 possessed the maximum
number of dominant genes for seeds per pod, on the other hand genotype Wanhar-2000
carried maximum number of recessive genes. The remaining parents falling in between two
were intermediate for distribution of the genes.
100-seed weight (g)
Diallel analysis for 100-seed weight (Table.20) revealed highly significant (P≤
0.01) mean squares due to additive (a) and dominant (b) genetic effects. The overall
dominance component b was smaller than a component indicated the pre-dominant role of
additive genetic effects. The item b1 and b2 were highly significant (P≤ 0.01) indicating the
presence of unidirectional dominance of the genes and the asymmetry of gene distribution
among parents respectively. The item b3 was highly significant which means that specific
gene interaction was present. Maternal effects (c) and reciprocal effects (d) were not
detected.
The estimation of genetic components of variation for 100-seed weight of genotype
(Table.21) revealed that both additive (2.12) and dominant component (1.48) of variation
were significant. However D component was greater than H1 indicated additive genes
controlling were predominating.
69
Table 20. Mean squares and degree of freedom for the analysis of variance of 6 ×
6 diallel for 100-seed weight (g) in chickpea.
ITEMS d.f MS F Ratio a 5 14.040 1546.880** b 15 1.926 209.822** b
1 1 4.685 1468.166**
b2 5 0.931 227.392**
b3 9 2.172 171.453**
c 5 0.010 0.807 d 10 0.011 1.039 Blocks 2 0.019 B × a 10 0.009 B × b 30 0.009 B × b
1 2 0.003
B × b2 10 0.004
B × b3 18 0.013
B × c 10 0.013 B × d 20 0.011 Bl.Int. 70 0.010 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
70
Table 21. Estimates of genetic components of variation for 100-Seed weight (g) in
chickpea.
COMPONENTS ESTIMATES
D 2.124* ±0.161
H1 1.482
* ±0.409
H2 1.277
* ±0.366
F 0.770NS
±0.394
h2 0.866
* ±0.246
E 0.003NS
±0.616
√H1/D 0.835
H2/4H
1 0.215
√4DH1+ F/ 4DH
1- F 1.554
h2 ns 0.7071
h2
bs 0.9968
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
71
Fig.9. Vr/Wr graph for 100-seed weight.
72
The estimates of H1 and H2 and H2/4H1 ratio 0.22 suggested the equal distribution
of positive and negative alleles among the parents. The value of F component of variation
(0.770) was positive and non-significant which indicated that dominant and recessive
alleles were statistically at par. Ratio of dominant to recessive genes (1.55) was greater
than 1 showing the dominant genes were more frequent than recessive genes. The net
dominance effects due to heterozygous loci (0.866) were also significant. The average
degree of dominance (0.835) was partial that was above the graphic origin intercept the
regression line (Fig.9) suggested that additive gene action with partial dominance was
present.
Environmental effects (E) were non-significant, indicated that no influence of
environment for the expression of the trait. The estimates of narrow and broad sense
heritability were high. Narrow sense heritability was (70.71%) of broad sense heritability.
Additive and dominant gene effects and high narrow-sense heritability were reported by
Chaturvedi et al. (1997), Bicer and Sakar (2008). He also reported additive gene effects
higher than dominant gene effects. Additive type of gene action was reported by Singh et
al. (1982), Singh et al. (1992), Kulkarni (2001) and Patil et al. (2006). Slightly variable
findings were published by Sandhu and Mandal (1989) and Kumar et al. (2001) who
revealed the predominance of additive variance for 100-Seed weight. Non-additive type of
gene action was reported by Singh and Bains (1982), Sarode et al. (2001) and Meena et al.
(2006). High heritability with low genetic advance was reported by Arshad et al. (2004).
Girase and Deshmukh (2000) observed duplicate type of gene action in the expression of
100-Seed weight.
Graphical analysis (Fig.9) for the character depicted that genotype Wanhar-2000
possessed the maximum number of dominant genes for 100-seed weight, on the other hand
genotype CM-98 carried maximum number of recessive genes. The remaining parents
falling in between two were intermediate for distribution of the genes.
73
Grain yield per plant (g)
Analysis of variance for grain yield per plant (Table.22) revealed highly significant
(P≤ 0.01) mean squares due to additive (a) and dominant (b) genetic effects. The overall
dominance component b was smaller than a component indicated the pre-dominant role of
additive genetic effects. The item b1 was significant (P≤ 0.05) and b2 was highly significant
(P≤ 0.01) indicating the presence of unidirectional dominance of the genes and asymmetry
of genes distribution among parents respectively. The item b3 was highly significant which
means that specific gene interaction was present. Maternal effects (c) and reciprocal effects
(d) were non-significant at (P≤ 0.05).
The estimation of genetic components of variation for grain yield per plant
(Table.23) revealed that both additive (19.15) and dominant component (21.02) of variation
were significant. However ‘D’ component was smaller than H1 indicated non-additive
types of gene action was present. The estimates of H1 and H2 and H2/4H1 ratio (0.19)
suggested the equal distribution of positive and negative alleles among the parents. The F
component of variation (14.69) was positive and non-significant which indicated that
dominant and recessive alleles were statistically at par. Ratio of dominant to recessive
genes (2.15) was greater than 1 indicated that dominant genes were more frequent than
recessive genes. The net dominance effects due to heterozygous loci (4.540) were non-
significant. The value of average degree of dominance (1.048) was greater than 1, intercept
below the graphic origin (Fig.10) the regression line, suggested over dominance type of
gene action for this trait.
Environmental effects (E) was non-significant, indicated that environmental effects
have little value for the expression of the character. The estimates of narrow and broad
sense heritability were high. Narrow sense heritability was (52.4%) of broad sense
heritability and dominance in nature. These results are in accordance with Biranvand et al.
(2008) who estimated genetic parameters D, H1 and H2, and indicated that additive
component of variance was less than non-additive components of variance, narrow sense
heritability was low and Wr/Vr graphs reflected over dominance for seed yield per plant.
Additive and dominance gene action was reported by Singh et al. (1992), Chaturvedi et al.
(1997), Sarode et al. (2001) and Beyran et al. (2008).
74
Table 22. Mean squares and degree of freedom for the analysis of variance of 6 × 6 diallel for grain yield per plant (g) in chickpea.
ITEMS d.f MS F Ratio a 5 83.694 168.648
**
b 15 24.777 52.456**
b
1 1 24.949 59.486
*
b2 5 22.002 67.873
**
b3 9 26.299 46.918
**
c 5 0.363 0.783 d 10 0.403 1.104 Blocks 2 0.046 B × a 10 0.469 B × b 30 0.472 B × b
1 2 0.419
B× b2 10 0.324
B × b3 18 0.560
B × c 10 0.464 B × d 20 0.364 Bl.Int. 70 0.444 Total 107
* = Significant at 5% level of probability ** = Significant at 1% level of probability
75
Table 23. Estimates of genetic components of variation for grain yield per plant in
chickpea.
COMPONENTS ESTIMATES
D 19.149* ±2.173
H1 21.022
* ±5.517
H2 16.229
* ±4.928
F 14.691* ±5.309
h2 4.540
NS ±3.317
E 0.144NS
±0.830
√H1/D 1.148
H2/4H
1 0.193
√4DH1+ F/ 4DH
1- F 2.155
h2 ns 0.5240
h2
bs 0.9836
* = Value is significant when it is exceeds 1.96 (t tabulated) after dividing it with its standard error NS = Non-significant
76
Fig.10. Vr/Wr graph for grain yield per plant.
77
Additive gene effects with high heritability were reported by Durga et al. (2007)
and Sreelakshmi et al. (2010). Non-additive type of gene action was reported by Kulkarni
(2001), Patil et al. (2006) and Meena et al. (2006).
Singh and Bains (1982), Malhotra et al. (1983), Tewari and Pandey (1986) and
Annigeri et al. (1996) observed both additive and non-additive genetic variance
significant for this trait. Predominance of epistatic gene action was observed by Patil et al.
(1998). Kumar et al. (2001) revealed the predominance of non-additive variance for this
trait. Husain et al. (1990) found that mode of inheritance appeared to be over-dominant for
grain yield per plant. Over-dominance in a positive sense and weak reciprocal differences
was reported by Martinez et al. (1978) for grain yield per plant. Girase and Deshmukh
(2000) observed grain yield predominantly under the control of dominance effect.
Graphical analysis (Fig.10) for the character depicted that genotype Balksar-2000
and Bittal-98 possessed maximum number of dominant genes for grain yield per plant, on
the other hand genotype CM-98 carried the maximum number of recessive genes. The
remaining parents falling in between two were intermediate for distribution of the genes.
2. Combining Ability
Good Combining Ability is the ability of a parent to produce better progeny when
combined with another parent. General combining ability (GCA) is mainly due to additive
gene action, while specific combining ability (SCA) is refers to the performance of two
particular genotypes in a specific cross, and is due to the non-additive types of gene
interactions. Statistically, GCA is the sum of the total effects of additive and additive ×
additive variances and SCA is the sum of total effects of dominance and dominance ×
dominance variances. Data obtained from the analysis of the diallel cross using combining
ability approach (Griffing, 1956) are presented below.
Number of days taken to flowering
Combining ability analysis for number of days taken to flowering revealed highly
significant mean squares (P≤ 0.01. Table.24) for general and specific combining abilities,
while mean squares due to reciprocal effects were non-significant.
78
Table 24. Mean squares and significances from the analysis of variance of
combining ability in 6×6 diallel cross of chickpea.
MEAN SQUARES CHARACTER GCA
(d.f =5) SCA (d.f =15)
Reciprocal (d.f =15)
Residual (d.f =70)
1 Number of days taken to flowering
39.95** 6.58** 0.23 0.20
2 Number of primary branches per plant
0.23** 0.01** 0.001** 0.0002
3 No. of secondary branches per plant
0.84** 0.02** 0.001 0.001
4 Plant height (cm) 133.22** 8.02** 7.39** 0.20
5 Number of days taken to maturity
105.03** 18.89** 0.54 0.23
6 Total weight of plant (g)
163.80** 9.20** 0.33** 0.06
7 Number of pods per plant
118.65** 25.85** 0.34 0.69
8 Number of seeds per pod
0.004** 0.002** 0.0003** 0.0001
9 100-seed weight (g) 4.68** 0.64** 0.004 0.003
10 Grain yield per plant (g)
27.90** 8.26** 0.13 0.15
11 Harvest index % 16.40** 9.32** 0.52 0.28
* = Significant at 5% probability level ** = Significant at 1% probability level NS = Non-significant
79
Table 25. Estimates of variation components, general (σ2g) & specific combing ability (σ2s), reciprocal effects (σ2r), error (σ2e) and GCA/SGA ratio in 6×6 diallel cross of chickpea
S.# Character (σ2g) (σ2s) (σ2r) (σ2e) GCA/
SCA
1 Number of days taken to flowering
3.38 3.31 0.01 0.20 1.02
2 Number of primary branches per plant
0.02 0.01 0.0006 0.0002 2.00
3 No. of secondary branches per plant
0.11 0.09 -3.8x10-05 0.001 1.22
4 Plant height (cm) 11.08 7.81 3.59 0.20 1.42
5 Number of days taken to maturity
8.73 18.66 0.15 0.23 0.46
6 Total weight of plant (g)
13.64 9.13 0.13 0.06 1.49
7 Number of pods per plant
9.82 25.15 -0.18 0.69 0.39
8 Number of seeds per pod
0.0003 0.002 9.96x10-5 0.0001 0.15
9 100-seed weight (g) 0.39 0.24 0.0001 0.003 1.62
10 Grain yield per plant (g)
2.31 8.11 -0.009 0.15 0.28
11 Harvest index % 1.34 9.04 0.11 0.28 0.15
80
The variance due to GCA was slightly greater than SCA variance (Table.25)
indicated that additive effects predominated in the expression of the character. The ratio of
the GCA/SCA variances (1.02) also supports the predominance of additive effects for
number of days taken to flowering. Singh et al. (1992) also reported pre-dominant gene
action under additive inheritance. Kulkarni (2001), Patil et al. (2006), Bicer and Sakar
(2008) and Bhardwaj et al. (2009) reported significant additive genetic effects for number
of days taken to flowering.
The combining ability and reciprocal effects are given in Table.26. The genotype
CM-98 was the best general combiner for number of days taken to flowering as it had
highest GCA value (1.204). Three parental genotypes AUG-786 (-0.833), Punjab-2000 (-
0.167) and Balksar-2000 (-0.333) was poor general combiner displayed negative GCA
effects. Nine of the fifteen direct crosses showed negative SCA effects and six showed
positive SCA effects. The best specific combiner was CM-98 × Punjab-2000 as it had
highest value 3.018. Reciprocal effects were negative in eight crosses while hybrid
Balksar-2000 × CM-98 was good specific combiner having the highest value (2.704).
Number of primary branches per plant
Mean squares for GCA, SCA and reciprocal effect were highly significant (P≤ 0.01,
Table.24) for number of primary branches per plant. Highly significant mean sum of
squares due to general and specific combining ability for number of primary branches per
plant was also recorded by Kulkarni (2001).
The variance due to GCA (0.02) was greater than SCA variance (0.01, Table.25) for
number of primary branches per plant, which suggested that additive genetic effects were
more prominent in controlling of this trait. These results are in agreement with observation
of Yadav et al. (1987), Bicer and Sakar (2008) who reported additive genetic control for
number of primary branches per plant. Katiyar et al. (1988) found both additive and non-
additive gene actions for this trait, with former being predominant for number of primary
branches per plant in chickpea genotypes.
CM-98 was the best general combiner (Table.26) for number of primary branches
per plant with maximum positive GCA effects (0.118) followed by Balksar-2000 (0.033)
while Punjab-2000 was the poorest general combiner with GCA -0.033 values for number
of primary branches per plant.
81
Table 26. General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea Genotypes and their crosses for number days taken to flowering.
S.E. (gi) = 0.118 S.E. (sij) = 0.268 S.E. (rij) = 0.316 Table 27. General combining ability (diagonal), Specific combining ability
(above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of primary branches per plant.
Parents CM-98 Bittal-98
AUG-786 Punjab- 2000
Wanhar-2000
Balksar-2000
CM-98 0.118 -0.004 -0.022 -0.012 0.096 0.101
Bittal-98 0.003 0.007 0.061 0.065 -0.044 -0.057
AUG-786 -0.099 -0.027 -0.002 0.118 0.011 -0.015
Punjab-2000 0.200 -0.013 0.028 -0.033 -0.100 -0.033
Wanhar-2000 -0.051 -0.023 -0.038 0.042 0.023 0.055
Balksar-2000 -0.171 0.008 -0.005 -0.035 -0.020 0.033
S.E. (gi) = 0.004 S.E. (sij) = 0.008 S.E. (rij) = 0.010
Parents CM-98 Bittal-98 AUG- 786
Punjab- 2000
Wanhar- 2000
Balksar- 2000
CM-98 1.204 -0.176 -1.593 3.018 -2.620 -0.398
Bittal-98 -1.852 0.001 -0.203 2.241 0.268 -2.676
AUG-786 -0.268 -0.167 -0.833 -1.009 0.518 0.241
Punjab-2000 0.287 -0.333 -0.167 -0.167 -1.704 -1.148
Wanhar-2000 -2.074 -0.333 0.500 0.001 0.001 2.546
Balksar-2000 2.704 0.002 0.001 -0.500 0.333 -0.333
82
Highest positive SCA effects (0.118) were recorded in the cross AUG-786 ×
Punjab-2000 as a good specific combiner while these effects were negative and highest (-
0.100) in the cross Punjab-2000 × Wanhar-2000. The hybrid Punjab-2000 × CM-98 (0.200)
was good specific combiner in reciprocal crosses. A total of ten crosses showed negative
reciprocal effects with highest value (-0.171) found in Balksar-2000 x CM-98 hybrid.
Number of secondary branches per plant
The parents differed significantly for GCA and SCA at P≤ 0.01 and reciprocal
effects were non-significant (Table.24) for number of secondary branches per plant. Similar
results were reported by Kulkarni (2001) and Patil et al. (2006). Dominant effects appeared
more pronounced for the trait manifestation (Table.25) as the variance for SCA (0.12) was
greater than GCA variance (0.07). The ratio of the GCA/SCA variances (0.58) was also
supported the predominance of additive effects for number of secondary branches per plant.
Yadav et al. (1987), Bicer and Sakar (2008) also found additive genetic effects to be
important in controlling this trait, while importance of additive and non-additive genetic
effects has been reported by Tewari and Pandey (1986) and Katiyar et al. (1988) for
number of secondary branches per plant.
The estimation of general combining ability, specific combining ability and
reciprocal effects are presented in Table.28. The parental line CM-98 had the highest
positive GCA effects of 0.294 indicating that this variety is good combiner for number of
secondary branches per plant while Bittal-98 is poor general combiner as it had the
negative GCA value of -0.017.
Nine of the direct cross combinations indicated positive SCA effects. Among them,
Wanhar-2000 × Balksar-2000 was the best SCA value = 0.366 while Punjab-2000 ×
Balksar-2000 was the poor specific combiner had negative SCA = -0.606 value. Positive
SCA effects were evident in 4 crosses as hybrid Punjab-2000 × CM-98 was good specific
combiner with highest positive (0.338).
83
Table 28. General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of secondary branches per plant.
Parents CM-98 Bittal-98 AUG-786 Punjab-2000
Wanhar-2000
Balksar-2000
CM-98 0.294 0.020 -0.200 0.061 -0.070 -0.035
Bittal-98 0.032 -0.017 0.028 0.180 -0.272 0.162
AUG-786 -0.257 -0.020 0.003 0.231 0.019 0.086
Punjab-2000 0.338 -0.003 -0.027 0.042 -0.183 -0.606
Wanhar-2000 -0.202 -0.045 -0.005 -0.027 0.0167 0.366
Balksar-2000 -0.205 -0.013 0.012 -0.013 0.0167 0.005
S.E. (gi) = 0.008 S.E. (sij) = 0.019 S.E. (rij) =0.022 Table 29. General combining ability (diagonal), Specific combining ability
(above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for plant height (cm).
Parents CM-98 Bittal-98 AUG-786 Punjab-2000
Wanhar-2000
Balksar-2000
CM-98 2.435 -1.407 3.370 -1.546 0.537 0.731
Bittal-98 0.491 0.001 -2.852 2.231 1.148 4.009
AUG-786 4.713 0.333 -2.833 0.676 0.426 -1.713
Punjab-2000 -4.537 2.833 -0.333 -1.000 -0.491 -1.130
Wanhar-2000 -0.620 3.500 -0.833 -1.333 -2.833 -0.713
Balksar-2000 -2.481 0.833 0.833 -2.000 -3.000 -1.000
S.E. (gi) = 0.119 S.E. (sij) = 0.270 S.E. (rij) = 0.318
84
Plant Height (cm)
The combining ability analysis indicated highly significant mean squares (P≤ 0.01,
Table.24) due to GCA, SCA and reciprocal effects. The mean sum of squares due to
general and specific combining ability highly significant also observed by Kulkarni (2001)
and Patil et al. (2006).
GCA variance was greater than SCA variance (Table.25) indicating relative
importance of additive genetic effects for plant height. These findings are in agreement
with observation of Bicer and Sakar (2008) and Bhardwaj et al. (2009) who reported
additive genetic control for this trait. Katiyar et al. (1988) found the importance of both
additive and non-additive gene actions, with former being predominant for plant height in
chickpea genotypes.
The estimation of general combining ability, specific combining ability and
reciprocal effects for plant height are presented in Table. 29. The parent CM-98 is good
general combiner for plant height as it had highest positive GCA effects of 2.435, while
AUG-2000 and Wanhar-2000 poor combiner, had the highest negative GCA value of -
2.833. Eight direct cross combinations showed positive SCA effects. Three crosses Bittal-
98 × Balksar-2000 (4.009), CM-98 × AUG-786 (3.370) and Bittal-98 × Punjab-2000
(2.231) displayed the higher positive SCA values where as the cross Bittal-98 × AUG-786
(-2.852) was the poor specific combiner for this trait. Reciprocal effects were higher SCA
value for AUG-786 × CM-98 cross indicated good specific combiner for plant height.
Number of days taken to maturity
Mean squares for number of days taken to maturity revealed significant (P≤ 0.01.
Table.24) differences to general and specific combining abilities, while variation due to
reciprocal effects was found non-significant. Kulkarni (2001) also observed highly
significant mean sum of squares due to general and specific combining ability.
The variance due to GCA was smaller than SCA variance (Table.25) indicating
more relative importance of non-additive effects for number of days taken to maturity.
Importance of non-additive genetic effects has also been reported by Meena et al. (2006)
for days taken to maturity.
85
Table 30. General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number days taken to maturity.
Parents CM-98 Bittal-98 AUG-786 Punjab- 2000
Wanhar-2000
Balksar-2000
CM-98 0.370 0.935 0.296 1.463 0.768 -2.426
Bittal-98 4.065 1.667 1.102 -0.231 0.907 -2.620
AUG-786 0.204 0.1667 0.001 3.296 -6.898 3.907
Punjab-2000 -1.463 0.001 -0.333 0.001 3.935 -3.093
Wanhar-2000 1.565 0.333 -0.500 -0.167 -0.333 2.380
Balksar-2000 -4.741 0.1667 0.667 -0.333 0.001 -0.167
S.E. (gi) = 0.127 S.E. (sij) = 0.289 S.E. (rij) = 0.340 Table 31. General combining ability (diagonal), Specific combining ability
(above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for total weight of plant (g).
Parents CM-98 Bittal-98 AUG-786 Punjab-2000
Wanhar-2000
Balksar-2000
CM-98 2.095 0.985 -0.943 -0.259 -1.054 -2.015
Bittal-98 2.679 -0.050 0.091 -1.026 -0.654 2.185
AUG-786 2.856 0.133 0.050 3.580 1.602 -4.559
Punjab-2000 0.940 0.001 0.050 0.033 -0.481 0.557
Wanhar-2000 -1.949 -0.083 0.067 0.033 -0.300 2.580
Balksar-2000 -6.621 0.450 0.167 0.433 0.900 -1.067
S.E. (gi) = 0.065 S.E. (sij) = 0.148 S.E. (rij) = 0.175
86
Singh et al.1992), Kabeta (2006), Anbessa et al. (2006) Bicer and Sakar (2008)
Biranvand (2008) and Bhardwaj et al. (2009) reported additive and non-dominant gene
effects were important in the inheritance of the trait, while Patil et al. (2006) observed
variance due to general and specific combining abilities was non-significant.
General and specific combining ability effects for days taken to maturity are
presented in Table.30. The parental line Bittal-98 (1.667) was the best combiner for days
taken to maturity. In contrast genotype Wanhar-2000 was the poor combiner for this trait as
it had the highest negative GCA value (-0.333). Four of six parental genotypes, Bittal-98
(1.667), CM-98 (0.370), AUG-786 (0.001) and Punjab-2000 (0.001) showed positive GCA
effects. Among the direct crosses 10 displayed positive SCA effects. Three hybrids are
good specific combiner with high positive SCA values were AUG-2000 × Balksar-2000
(3.907), AUG-786 × Punjab-2000 (3.296), and Wanhar-2000 × Balksar-2000 (2.380).
AUG-2000 × Wanhar-2000 (-6.898) had lowest SCA value indicating that this cross is poor
specific combiner. Positive and highest SCA effects (4.065) were recorded in reciprocal
cross Bittal-98 × CM-98 indicating good specific combing ability for number of days taken
to maturity.
Total weight of plant (g)
Analysis of variance for GCA, SCA and reciprocal effects were significant at P≤
0.01 (Table.24) for total weight of plant. Highly significant mean squares due to GCA and
SCA were also reported by Kulkarni (2001) for total weight of plant.
It was revealed in Table.25 that GCA variance (13.64) was greater than SCA
variance (9.13) for total weight of plant, which suggested that additive genetic effects were
more prominent in controlling the genetic of this trait. Singh and Bains (1985), Arshad et
al. (2004) Venkatraman (2007) and Bhardwaj et al. (2009) were also observed GCA
variances significant and of high magnitude, while specific combining ability (SCA)
variances were non-significant. Non-additive type of gene action was reported by Meena et
al. (2006) on biomass. Over dominant type gene action was indicated by Hussain et al.
(1990) for this trait. Tewari and Pandey (1986) revealed significant role of additive and
non-additive gene actions for total weight of plant.
CM-98 was the best combiner (Table.31) for total weight of plant with maximum
positive GCA effects (2.095) fallowed by AUG-786 effects (0.050) while Balksar-2000
87
showed highest negative GCA effects (-1.067) is the poor general combiner for total weight
of plant. Highest positive SCA effects (3.580) were recorded in the cross AUG-786 ×
Punjab-2000 while these effects were negative and highest (-4.559) in the cross AUG-786
× Balksar-2000. Positive SCA effects were evident in 12 cross combinations, with highest
value (2.856) observed in hybrid AUG-786 × CM-98 in reciprocal crosses.
Number of pods per plant
Mean squares for GCA and SCA were significant (P≤ 0.01, Table.24) for number of
pods per plant, while variation due to reciprocal effects was found non-significant. Similar
results were reported by Kulkarni (2001) and Patil et al. (2006).
The variance due to GCA was smaller than SCA variance and GCA/SCA ratio
(Table.25) suggested that the non-additive effects were more prominent in controlling of
genetics for number of pods per plant. Yadav et al. (1987), Bicer and Sakar (2008) and
Bhardwaj et al. (2009) observed additive genetic effects to be important in controlling this
trait, while importance of additive and non-additive genetic effects for number of pods per
plant has been reported by Tewari and Pandey (1986), Chaturvedi (1997) and Katiyar et al.
(1988). Hussain et al. (1990) assessed general and specific combining abilities and found
that mode of inheritance appeared to be over-dominant for number of pods per plant.
General and specific combining ability effects for number of pods per plant are
presented in Table.32. CM-98 was the best general combiner for number of pods per plant
fallowed by Bittal-98 (0.667), Balksar-2000 (0.667) and Punjab-2000 (0.333) indicated
positive GCA effects. The genotype, AUG-786 was poor combiner had the negative GCA
value of -0.500 fallowed by Wanhar-2000 had value of -0.167.
Among the direct crosses 8 displayed positive SCA effects. Three hybrids with high
positive SCA values were Bittal-98 × Wanhar-2000 (5.685), Punjab-2000 × Balksar-2000
(4.491), AUG-786 × Punjab-2000 (4.435) and Wanhar-2000 × Balksar-2000 (4.296) are
good specific combiner while AUG-2000 × Balksar-2000 (-5.648) had lowest SCA value.
Positive and highest SCA effects (2.565) were recorded in hybrid Bittal-98 × CM-98 for
reciprocal crosses.
88
Table 32. General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of pods per plant.
Parents CM-98 Bittal-98 AUG-786 Punjab-2000
Wanhar-2000
Balksar-2000
CM-98 2.204 -1.120 2.769 0.074 -2.4537 0.157
Bittal-98 2.565 0.667 1.741 -0.120 5.685 -1.704
AUG-786 0.510 -0.167 -0.500 4.435 -3.259 -5.648
Punjab-2000 0.704 -0.333 -0.500 0.333 -2.454 4.491
Wanhar-2000 -6.102 0.333 0.167 -0.500 -0.167 4.296
Balksar-2000 0.120 -0.167 0.001 0.001 -0.667 0.667
S.E. (gi) = 0.219 S.E. (sij) = 0.500 S.E. (rij) = 0.588
Table 33. General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for number of seeds per pod.
Parents CM-98 Bittal-98 AUG-786 Punjab- 2000
Wanhar-2000
Balksar-2000
CM-98 -0.015 -0.023 0.017 -0.016 0.018 -0.028
Bittal-98 -0.002 -0.013 0.001 -0.033 0.054 -0.019
AUG-786 -0.018 0.000 0.001 0.016 -0.065 0.010
Punjab-2000 -0.005 0.001 -0.007 0.002 -0.011 0.064
Wanhar-2000 0.035 0.022 -0.007 0.015 -0.005 -0.006
Balksar-2000 0.005 -0.002 -0.003 -0.033 -0.017 -0.007
S.E. (gi) = 0.003 S.E. (sij) = 0.007 S.E. (rij) = 0.008
89
Number of seeds per pod
Analysis of variance for combining ability revealed significant (P≤ 0.01, Table.24)
means squares due to GCA, SCA and reciprocal effects for number of seeds per pod.
Similar results were reported by Kulkarni (2001).
Non-additive genetic effects for number of seeds per pod (Table.25) appeared more
pronounced for the character manifestation as SCA variance (0.002) was greater than GCA
variance (0.0003). Chaturvedi (1997) revealed both additive as well as non-additive
components of variance were important for number of seeds per pod while Bicer and Sakar
(2008) and Bhardwaj et al. (2009) were found significant additive genetic variance for for
number of seeds per pod.
Combining ability effects (Table.33) indicated that only two parents, Punjab-2000
(0.002) and AUG-786 (0.001) were good general combiner while CM-98 (GCA= -0.015)
was the poor general combiner for number of seeds per plant. Positive SCA effects were
recorded in 7 cross combinations and were maximum in Punjab-2000 × Balksar-2000
(0.064). The poor specific combiner was AUG-786 × Wanhar-2000 as it had the negative
SCA effects (-0.065). In reciprocal cross combinations SCA effects were negative in 9
crosses with highest value (-0.033) observed in Balksar-2000 × Punjab-2000 hybrid.
100-seed weight (g)
Analysis of variance for 100-seed weight revealed highly significant (P≤ 0.01
Table.24) mean squares due to GCA and SCA effects. Kulkarni (2001) was also reported
highly significant mean squares due to GCA and SCA for 100-seed weight.
SCA variance (0.24) was smaller than GCA variance (0.39) indicated that additive genetic
effects were more important for 100-seed weight (Table.25). Bicer and Sakar (2008) and
Bhardwaj et al. (2009) also reported additive genetic effects to be important in controlling
this trait, while importance of additive and non-additive genetic effects for 100-seed weight
observed by Chaturvedi (1997) and Katiyar et al. (1988). Patil et al. (2006) reported
variances due to general and specific combining abilities were significant.
The estimation of general combining ability, specific combining ability and
reciprocal effects is given in Table.34. The parents have shown differences in GCA effects
for 100-seed weight. The parent CM-98 (0.398) was the best combiner, had highest
90
Table 34. General combining ability (diagonal), Specific combining ability (above diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for 100-seed weight (g).
Parents CM-98 Bittal-98 AUG-786 Punjab-2000
Wanhar-2000
Balksar-2000
CM-98 0.398 0.620 0.225 0.186 0.004 -0.962
Bittal-98 0.533 -0.012 -0.611 -0.017 0.012 0.270
AUG-786 0.324 -0.008 -0.020 0.518 -0.310 0.192
Punjab-2000 0.098 0.001 0.162 -0.005 0.348 -0.193
Wanhar-2000 -0.187 -0.025 0.002 -0.002 0.003 1.117
Balksar-2000 -1.165 -0.002 0.002 -0.002 -0.005 0.005
S.E. (gi) = 0.015 S.E. (sij) = 0.035 S.E. (rij) = 0.041 Table 35. General combining ability (diagonal), Specific combining ability (above
diagonal) and reciprocal (below diagonal) effects of 6 chickpea genotypes and their crosses for grain yield per plant (g).
S.E. (gi) = 0.101 S.E. (sij) = 0.231 S.E. (rij) = 0.272
Parents CM-98 Bittal-98 AUG-786 Punjab- 2000
Wanhar-2000
Balksar-2000
CM-98 1.330 0.061 0.706 -0.025 -1.024 -2.354
Bittal-98 1.752 0.187 -0.150 -0.557 3.116 -0.689
AUG-786 0.481 0.112 -0.560 2.945 -1.945 -1.347
Punjab-2000 0.051 -0.133 -0.083 0.168 -0.675 2.035
Wanhar-2000 -1.914 0.513 0.108 -0.077 -0.157 3.126
Balksar-2000 -1.700 -0.190 -0.063 -0.065 -0.437 0.140
91
positive GCA effect, while AUG-786 possessing highest negative GCA effects (-0.02), was
the poor combiner. Wanhar-2000 × Balksar-2000 was the best specific combination with
the highest positive SCA effect of 1.117 and CM-98 × Balksar-2000 was the poor specific
cross with the highest negative SCA effect of -0.962. In reciprocal crosses SCA effects
was positive and highest in the cross Bittal-98 × CM-98 (0.533).
Grain yield per plant (g)
Mean squares for GCA and SCA were highly significant (P≤ 0.01 Table.24) and
reciprocal effects were non-significant for grain yield per plant. Highly significant mean
squares due to GCA and SCA were also reported by Kulkarni (2001) for grain yield per
plant.
The higher SCA variance (8.11) relative to GCA variance (2.31) indicated that non-
additive genetic effects were more important in the trait expression (Table.25). Yadav et al.
(1987) and Bhardwaj et al. (2009) observed additive variance was significant. Importance
of additive and non-additive genetic effects for grain yield per plant has been observed by
Tewari and Pandey (1986), Chaturvedi (1997) and Katiyar et al. (1988). Hussain et al.
(1990) assessed general and specific combining abilities and found over-dominant type of
gene action for grain yield per plant. Patil et al. (2006) reported significant variances due to
general and specific combining abilities.
The estimation of the general combining, specific combining ability and reciprocal
effects is given in Table.35. CM-98 was the best general combiner for grain yield per plant
with maximum positive GCA effects (1.330) followed by Bittal-98 (0.187) while AUG-786
showed highest negative GCA effect (-0.560) was the poor general combiner for this trait.
Highest positive SCA effects were estimated in the crosses Wanhar-2000 × Balksar-2000
(3.126) and Bittal-98 × Wanhar-2000 (3.116) while these effects were highest and negative
(-2.354) in the cross CM-98 × Balksar-2000.A total of 9 crosses recorded negative SCA
effects with highest negative value (-1.914) in Wanhar-2000 × CM-98 hybrid for reciprocal
crosses.
92
Table 36. General combining ability (diagonal), Specific combining ability
(above diagonal) and reciprocal (below diagonal) effects of 6 chickpea
genotypes and their crosses for harvest index (%).
Parents CM-98 Bittal-98 AUG-786 Punjab-2000
Wanhar-2000
Balksar-2000
CM-98 0.399 -0.451 1.402 0.172 -0.619 -1.892
Bittal-98 0.648 0.249 -0.248 -0.104 4.392 -2.268
AUG-786 -0.988 0.059 -0.695 -0.248 -0.104 4.392
Punjab-2000 -0.489 -0.166 -0.129 0.178 -0.613 2.372
Wanhar-2000 -1.358 0.706 0.097 -0.109 -0.038 2.620
Balksar-2000 1.788 -0.550 -0.183 -0.383 -1.197 0.903
S.E. (gi) = 0.141 S.E. (sij) = 0.321 S.E. (rij) = 0.378
93
Harvest index (%)
Analysis of variance for harvest index revealed significant (P≤ 0.01 Table.24) mean
squares due to GCA and SCA effects. Highly significant mean squares due to GCA and
SCA for harvest index was also reported by Kulkarni (2001).
SCA variance (9.04) was greater than GCA variance (1.34) indicating the
predominance of non-additive genetic control for harvest index (Table.25). Importance of
additive and non-additive genetic effects has been observed by Chaturvedi (1997) for
harvest index. Patil et al. (2006) reported Variances due to general and specific combining
abilities were significant. Venkatraman (2007) observed GCA variances significant and of
high magnitude, while specific combining ability (SCA) variances were non-significant,
indicating the predominant role of additive gene action on harvest index.
The estimation of general combining ability, specific combining ability and
reciprocal effects is given in Table.36. The parents have shown differences in GCA effects
for harvest index. The parent Balksar-2000 (0.903) had highest positive GCA effect is the
best combiner, whilst AUG-786 possessing highest negative GCA effects (-0.695) is the
poor general combiner. AUG-786 × Balksar-2000 and Bittal-98 × Wanhar-2000 were the
best specific cross combinations with the highest positive SCA effects of 4.392 and Bittal-
98 × Balksar-2000 was the poor specific combiner with the highest negative SCA effect of
-2.268. Reciprocal effects was positive and highest SCA in the cross Balksar-2000 × CM-
98 (1.788) while 10 crosses were shown negative SCA effects.
3. Molecular Studies
Gel pictures showing amplifications in parents and hybrids are presented in figures
11-17. The six parents and their hybrids (21) were carefully studied on the basis of bands.
The level of polymorphism was different with different primers. A total of 28 primers were
used in PCR. Out of 28 primers, 21 were amplified the genomic DNA in all the varieties,15
primers amplified polymorphic DNA bands among all the varieties and 6 produced similar
banding pattern. Approximately 25% of the total reactions could not be amplified genomic
DNA. This is due to contamination in reaction mixture may cause primer degeneration
resulted into complete failure of amplification. The amount of DNA used in the
amplification reaction is 5ng. In the PCR reactions some primers amplified very bright and
94
some faint DNA bands. The extremely bright bands may be produced by the amplification
of high copy number sequences in the genomic DNA. Faint bands may be produced due to
mismatch of primers at complimentary sites, resulting in a poor amplification of genomic
DNA region. After scoring the bands, similarity matrix was developed (Table.37) after
multivariate analysis using Nei & Li’s (1979) coefficients. The similarity coefficients were
used to develop a dendrogram by UPGMA analysis in order to determine the cluster of the
varieties.
The cluster of six chickpea varieties and their hybrids revealed associations in the
experimental materials as shown in dendrogram. Chickpea genotypes were classified into
six main clusters namely a, b, c, d, e and f. The cluster ‘a’ was genetically more diverse to
all others. The cluster ‘c’ was further sub divided into four sub clusters. The two parents
Wanhar-2000 and Balksar-2000 were fall in one cluster. The parents Punjab-2000 and
Bittal-98 was also in same cluster. The results of our findings are indicative of their genetic
association. To increase more diversity breeders should use parents with diverse origin. It
will also help to choose parents for further breeding program.
Williams et al. (1990), Rakesh et al. (2002), Iruela et al. (2002), Souframanien and
Gopalakrishna (2004), Cingilli and Akcin (2005), Rao et al. (2007) and Talebi et al.
(2009) studied the polymorphism in chickpea using RAPD primers. Welsh and McCielland
(1990) studied the polymorphism in five species of Staphylococcus, eleven strains of
Staphylococcus pyogenes and three varieties of Oryza sativa, using RAPD primers.
95
Table37. Similarity matrix for Nei and Li's Original Measures of Genetic Identity and Genetic distance of 21 Chickpea genotypes
==================================================================================================================================================================================================
Pop 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
Bal× Wan× Wan× B-98× Pb-98× A-786× B-98 Wan× B-98 CM98 A-786 CM98 Balksar A-786 Balksar Pb.2000 Wanhar Balksar CM-98 AUG-786 Bittal-98
Bittal-98 Pb-2000 B-98 CM-98 Balksar Pb-2000 A-786 Balkas Pb-2000 Wanhar CM-98 Pb2000 A-786 Wanhar CM-98
==============================================================================================================================================
1 **** 0.7021 0.5957 0.6383 0.6170 0.6809 0.5957 0.6170 0.7021 0.6596 0.6383 0.7021 0.6383 0.6596 0.6809 0.6170 0.6596 0.7234 0.7021 0.7660 0.7234
2 0.3536 **** 0.7660 0.7234 0.6596 0.6809 0.5532 0.6170 0.7021 0.7447 0.7234 0.6596 0.5957 0.7447 0.6383 0.5745 0.7021 0.7234 0.6596 0.8085 0.6809
3 0.5179 0.2666 **** 0.7872 0.6809 0.7872 0.7447 0.6809 0.6809 0.7234 0.7447 0.7234 0.6170 0.6383 0.5745 0.5532 0.7660 0.7872 0.7234 0.6596 0.5319
4 0.4490 0.3238 0.2392 **** 0.7660 0.7021 0.7447 0.6383 0.6809 0.7660 0.7447 0.6383 0.5319 0.6809 0.6596 0.6809 0.7234 0.7872 0.8511 0.7021 0.6170
5 0.4829 0.4162 0.3844 0.2666 **** 0.7234 0.7660 0.5319 0.7447 0.7872 0.6809 0.7021 0.6809 0.7447 0.6383 0.6596 0.7872 0.8085 0.8298 0.6809 0.6809
6 0.3844 0.3844 0.2392 0.3536 0.3238 **** 0.7872 0.6809 0.8085 0.7660 0.7872 0.8511 0.7447 0.8085 0.7021 0.6383 0.8085 0.8723 0.8085 0.6596 0.6170
7 0.5179 0.5921 0.2948 0.2948 0.2666 0.2392 **** 0.5532 0.7234 0.7234 0.7021 0.7234 0.6170 0.7234 0.6170 0.6809 0.7660 0.7872 0.7660 0.6170 0.6170
8 0.4829 0.4829 0.3844 0.4490 0.6313 0.3844 0.5921 **** 0.7447 0.7021 0.7660 0.7021 0.7234 0.6170 0.6809 0.7447 0.6596 0.6809 0.6170 0.5532 0.5532
9 0.3536 0.3536 0.3844 0.3844 0.2948 0.2126 0.3238 0.2948 **** 0.9149 0.8936 0.8723 0.8085 0.8298 0.7660 0.7872 0.8723 0.8511 0.7872 0.7234 0.7234
10 0.4162 0.2948 0.3238 0.2666 0.2392 0.2666 0.3238 0.3536 0.0889 **** 0.8511 0.7872 0.7234 0.7872 0.6809 0.7872 0.9149 0.8936 0.8298 0.6809 0.7234
11 0.4490 0.3238 0.2948 0.2948 0.3844 0.2392 0.3536 0.2666 0.1125 0.1613 **** 0.8936 0.7872 0.8085 0.7021 0.7660 0.7660 0.8298 0.7660 0.6596 0.7021
12 0.3536 0.4162 0.3238 0.4490 0.3536 0.1613 0.3238 0.3536 0.1366 0.2392 0.1125 **** 0.8936 0.7872 0.7234 0.7447 0.7447 0.8511 0.7447 0.6383 0.6809
13 0.4490 0.5179 0.4829 0.6313 0.3844 0.2948 0.4829 0.3238 0.2126 0.3238 0.2392 0.1125 **** 0.7234 0.6596 0.7660 0.6809 0.7447 0.6809 0.5745 0.6596
14 0.4162 0.2948 0.4490 0.3844 0.2948 0.2126 0.3238 0.4829 0.1866 0.2392 0.2126 0.2392 0.3238 **** 0.7234 0.7447 0.7447 0.7660 0.7447 0.7234 0.7234
15 0.3844 0.4490 0.5543 0.4162 0.4490 0.3536 0.4829 0.3844 0.2666 0.3844 0.3536 0.3238 0.4162 0.3238 **** 0.7234 0.6383 0.7021 0.6809 0.6596 0.5745
16 0.4829 0.5543 0.5921 0.3844 0.4162 0.4490 0.3844 0.2948 0.2392 0.2392 0.2666 0.2948 0.2666 0.2948 0.3238 **** 0.7447 0.7660 0.7447 0.6383 0.7660
17 0.4162 0.3536 0.2666 0.3238 0.2392 0.2126 0.2666 0.4162 0.1366 0.0889 0.2666 0.2948 0.3844 0.2948 0.4490 0.2948 **** 0.8936 0.8298 0.7234 0.7234
18 0.3238 0.3238 0.2392 0.2392 0.2126 0.1366 0.2392 0.3844 0.1613 0.1125 0.1866 0.1613 0.2948 0.2666 0.3536 0.2666 0.1125 **** 0.8936 0.7021 0.7447
19 0.3536 0.4162 0.3238 0.1613 0.1866 0.2126 0.2666 0.4829 0.2392 0.1866 0.2666 0.2948 0.3844 0.2948 0.3844 0.2948 0.1866 0.1125 **** 0.7660 0.7660
20 0.2666 0.2126 0.4162 0.3536 0.3844 0.4162 0.4829 0.5921 0.3238 0.3844 0.4162 0.4490 0.5543 0.3238 0.4162 0.4490 0.3238 0.3536 0.2666 **** 0.7872
21 0.3238 0.3844 0.6313 0.4829 0.3844 0.4829 0.4829 0.5921 0.3238 0.3238 0.3536 0.3844 0.4162 0.3238 0.5543 0.2666 0.3238 0.2948 0.2666 0.2392 ****
===============================================================================================================================================
Nei's genetic identity (above diagonal) and genetic distance (below diagonal).
96
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 M
Fig.11: Amplification profile of 21 Chickpea genotypes with primer GLD-11. M
is a 1Kb ladder. Balksar-2000 × Bittal-98 (1), Wanhar-2000 × Punjab-2000 (2),
Wanhar-2000 × Bittal-98 (4), Punjab-2000 × Balksar-2000 (5), AUG-786 ×
Punjab-2000 (6), Bittal-98 × AUG-786 (7), Wanhar-2000 × Balksar-2000 (8),
Bittal-98 × Punjab-2000 (9), CM-98 × Wanhar-2000 (10), AUG-786 × CM-98
(11), CM-98 × Punjab-2000 (12), Balksar-2000 × AUG-786 (13), AUG-786 ×
Wanhar-2000 (14), Balksar-2000 × CM-98 (15), Punjab-2000 (16), Wanhar-2000
(17), Balksar-2000 (18), CM-98 (19), AUG-786 (20), Bittal-98 (21).
97
2 3 4 9 10 11 M
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 M
Fig. 12. Amplification profile of 21 Chickpea genotypes with primer GLA-01.
M is a 1Kb ladder. Balksar-2000 × Bittal-98 (1), Wanhar-2000 × Punjab-2000
(2), Wanhar-2000 × Bittal-98 (4), Punjab-2000 × Balksar-2000 (5), AUG-786
× Punjab-2000 (6), Bittal-98 × AUG-786 (7), Wanhar-2000 × Balksar-2000
(8), Bittal-98 × Punjab-2000 (9), CM-98 × Wanhar-2000 (10), AUG-786 ×
CM-98 (11), CM-98 × Punjab-2000 (12), Balksar-2000 × AUG-786 (13),
AUG-786 × Wanhar-2000 (14), Balksar-2000 × CM-98 (15), Punjab-2000 (16),
Wanhar-2000 (17), Balksar-2000 (18), CM-98 (19), AUG-786 (20), Bittal-98
(21).
98
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21M
Fig. 13: Amplification profile of 21 Chickpea genotypes with primer GLD-01. M
is a 1Kb ladder. Balksar-2000 × Bittal-98 (1), Wanhar-2000 × Punjab-2000 (2),
Wanhar-2000 × Bittal-98 (4), Punjab-2000 × Balksar-2000 (5), AUG-786 ×
Punjab-2000 (6), Bittal-98 × AUG-786 (7), Wanhar-2000 × Balksar-2000 (8),
Bittal-98 × Punjab-2000 (9), CM-98 × Wanhar-2000 (10), AUG-786 × CM-98
(11), CM-98 × Punjab-2000 (12), Balksar-2000 × AUG-786 (13), AUG-786 ×
Wanhar-2000 (14), Balksar-2000 × CM-98 (15), Punjab-2000 (16), Wanhar-2000
(17), Balksar-2000 (18), CM-98 (19), AUG-786 (20), Bittal-98 (21).
99
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 M
Fig.14. Amplification profile of 21 Chickpea genotypes with primer GLB-14. M
is a 1Kb ladder. Balksar-2000 × Bittal-98 (1), Wanhar-2000 × Punjab-2000 (2),
Wanhar-2000 × Bittal-98 (4), Punjab-2000 × Balksar-2000 (5), AUG-786 ×
Punjab-2000 (6), Bittal-98 × AUG-786 (7), Wanhar-2000 × Balksar-2000 (8),
Bittal-98 × Punjab-2000 (9), CM-98 × Wanhar-2000 (10), AUG-786 × CM-98
(11), CM-98 × Punjab-2000 (12), Balksar-2000 × AUG-786 (13), AUG-786 ×
Wanhar-2000 (14), Balksar-2000 × CM-98 (15), Punjab-2000 (16), Wanhar-
2000 (17), Balksar-2000 (18), CM-98 (19), AUG-786 (20), Bittal-98 (21).
100
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 M
Fig.15. Amplification profile of 21 Chickpea genotypes with primer GLC-20.
M is a 1Kb ladder. Balksar-2000 × Bittal-98 (1), Wanhar-2000 × Punjab-
2000 (2), Wanhar-2000 × Bittal-98 (4), Punjab-2000 × Balksar-2000 (5),
AUG-786 × Punjab-2000 (6), Bittal-98 × AUG-786 (7), Wanhar-2000 ×
Balksar-2000 (8), Bittal-98 × Punjab-2000 (9), CM-98 × Wanhar-2000 (10),
AUG-786 × CM-98 (11), CM-98 × Punjab-2000 (12), Balksar-2000 × AUG-
786 (13), AUG-786 × Wanhar-2000 (14), Balksar-2000 × CM-98 (15),
Punjab-2000 (16), Wanhar-2000 (17), Balksar-2000 (18), CM-98 (19), AUG-
786 (20), Bittal-98 (21).
101
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 M
Fig.16. Amplification profile of 21 Chickpea genotypes with primer GLA-
09. M is a 1Kb ladder. Balksar-2000 × Bittal-98 (1), Wanhar-2000 ×
Punjab-2000 (2), Wanhar-2000 × Bittal-98 (4), Punjab-2000 × Balksar-
2000 (5), AUG-786 × Punjab-2000 (6), Bittal-98 × AUG-786 (7), Wanhar-
2000 × Balksar-2000 (8), Bittal-98 × Punjab-2000 (9), CM-98 × Wanhar-
2000 (10), AUG-786 × CM-98 (11), CM-98 × Punjab-2000 (12), Balksar-
2000 × AUG-786 (13), AUG-786 × Wanhar-2000 (14), Balksar-2000 ×
CM-98 (15), Punjab-2000 (16), Wanhar-2000 (17), Balksar-2000 (18),
CM-98 (19), AUG-786 (20), Bittal-98 (21).
102
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 M
Fig.17. Amplification profile of 21 Chickpea genotypes with primer
GLA-02. M is a 1Kb ladder. Balksar-2000 × Bittal-98 (1), Wanhar-2000
× Punjab-2000 (2), Wanhar-2000 × Bittal-98 (4), Punjab-2000 ×
Balksar-2000 (5), AUG-786 × Punjab-2000 (6), Bittal-98 × AUG-786
(7), Wanhar-2000 × Balksar-2000 (8), Bittal-98 × Punjab-2000 (9), CM-
98 × Wanhar-2000 (10), AUG-786 × CM-98 (11), CM-98 × Punjab-2000
(12), Balksar-2000 × AUG-786 (13), AUG-786 × Wanhar-2000 (14),
Balksar-2000 × CM-98 (15), Punjab-2000 (16), Wanhar-2000 (17),
Balksar-2000 (18), CM-98 (19), AUG-786 (20), Bittal-98 (21).
.
103
104
4. Inheritance of resistance to chickpea wilt.
All six genotypes CM-98, AUG-786, Bittal-98, Balksar-2000, Wanhar-2000 and
Punjab-2000 were resistant. The crosses of all six genotypes with AUG-424 were
moderately susceptible in F1 progenies responded to inoculation by the formation of
necrotic local lesions on primary leaves and later on no distortion or stunting of plants was
observed. F2s segregated in a ratio of 1 susceptible 2 moderately susceptible : 1 resistant;
and F3 progenies, 1 susceptible : 2 segregating : 1 resistant ( Table.38). The data were
consisting with segregation of a single gene with the recessive allele conferring resistance
to race 1 of the wilt fungus. The heterozygous populations of all cross combinations in F3
generation also advocated a close fit of actual to the expected distribution for a 1:2:1 ratio
indicating resistance was controlled by a single gene pair in crosses under study. There was
an excess of resistant progenies in the F3 generation from Wanhar-2000 × AUG-424 and
Punjab-2000 × AUG-2000 that could have resulted from a failure of occurrence. Some
crosses did not give good fits to the expected ratios for simple inheritance.
Ayyar and Iyer (1936), Kumar and Haware (1982), Upadhyaya et al. (1983), Kumar
(1998), Shah et al. (2009) and Ahmad et al. (2010) observed mono genic inheritance for
wilt inheritance. Haware and Nene (1982) reported four races of F. oxysporum, only a few
reports are available on race 2 of Fusarium wilt. Pathak et al. (1975) reported that under
field condition the resistance to race 2 was controlled by a single recessive gene, whereas
preliminary finding of Gumber et al. (1995) suggested the involvement of two genes.
Navas-Cortés et al. (2000) studied the losses due to development of Fusarium wilt
epidemics.
105
Table 38. Number of resistant and susceptible plants or progenies in F1, F2 and F3
generations of the 6 crosses infested with Fusarium oxysporum L.
Cross combination
Number of plants observed Generation
Total Resistant Segregating Susceptible
Expected ratio
d.f X2 P-value
CM-98 × AUG-424
F1 14 14
F2 367 85 201 81 1:2:1 2 3.419 0.70-0.50
F3 609 155 315 139 1:2:1 2 1.569 0.50-0.30
Bittal-98 × AUG-424
F1 17 17
F2 396 81 208 107 1:2:1 2 4.424 0.50-0.30
F3 428 125 201 102 1:2:1 2 4.052 0.30-0.20
AUG-786 × AUG-424
F1 11 11
F2 81 27 39 15 1:2:1 2 3.725 0.70-0.50
F3 342 99 147 96 1:2:1 2 6.842 0.10-0.05 Punjab-2000 × AUG-424
F1 13 13
F2 508 124 265 119 1:2:1 2 1.051 0.70-0.50
F3 350 108 167 75 1:2:1 2 7.006 0.10-0.05 Wanhar-2000 × AUG-424
F1 8 8
F2 757 193 388 176 1:2:1 2 1.243 0.50-0.30
F3 989 271 501 217 1:2:1 2 6.075 0.10-0.05 Balksar-2000 × AUG-424
F1 9 9
F2 145 29 83 33 1:2:1 2 4.736 0.50-0.30
F3 343 104 156 83 1:2:1 2 5.360 0.10-0.05
Total 1401 2587 1243 1:2:1 2 9.967 0.30-0.20
106
Chapter-5
SUMMARY
The objective of the present study was to identify parental genotypes or crosses that
can be used in future breeding programs to develop new high yielding varieties of chickpea
with a wide range of variability and resistant to wilt disease. Gene action and combining
ability estimates were made by using diallel analysis. Data pertaining to various traits of six
chickpea genotypes i.e. CM-98, AUG-786, Bittal-98, Balksar-2000, Wanhar-2000 and
Punjab-2000 were analyzed following the method Mather and Jinks (1982) and Hayman
(1954). The traits studied included days taken to flowering, number of primary branches
per plant, number of secondary branches per plant, plant height, days taken to maturity,
total weight of plant, number of pods per plant, number of seeds per pod, 100-seed weight,
grain yield per plant, harvest index and wilt resistance.
A dendrogram was generated by using UPGMA analysis (Nei and Li’s, 1979) to
ascertain the extent of diversity among the parents and F1 crosses. Wilt inheritance was
estimated, six successful crosses among resistant and susceptible lines were manipulated to
raise F1, F2 and F3 generations to evaluate disease reaction and data were subjected to chi-
square test.
Scaling tests were used to test adequacy of the data for analysis using additive-
dominance model. The test showed that except harvest index, data were adequate or
partially adequate for the traits under study. Analysis of variance of diallel tables revealed
that additive (a) and dominance (b) effects were significant (P≤ 0.01) suggesting the
importance of both additive and dominance variation in the inheritance in all the traits. The
estimates of average degree of dominance showed partial dominance for number of days
taken to flowering, plant height, number of primary branches per plant, number of
secondary branches per plant, total weight of plant and 100-seed weight. Complete
dominance was exhibited for pods per plant and over-dominance observed for days taken to
maturity, number of seeds per pod, grain yield per plant and harvest index.
107
Additive effects appeared to be more important than the dominant effects for
number of days taken to flowering, number of primary branches per plant, number of
secondary branches per plant, plant height, total weight of plant and 100-seed weight
suggesting the importance of additive variation in the inheritance of these characters.
Moderate to high narrow-sense heritability was manifested for number of days
taken to flowering, number of primary branches per plant, number of secondary branches
per plant, plant height, total weight of plant and 100-seed weight due to additive gene
action. The remaining traits showing moderate to low heritability estimates (59.7 to 37.8%)
needed greater selection pressure through recurrent selection in early segregating
populations.
The mean squares due to GCA and SCA were highly significant for days taken to
flowering, number of primary branches per plant, number of secondary branches per plant,
plant height, days taken to maturity, total weight of plant, number of pods per plant,
number of seeds per pod, 100-seed weight, grain yield per plant and harvest index. GCA
effects suggested that CM-98 was the best general combiner for days taken to flowering,
number of primary branches per plant, number of secondary branches per plant, plant
height, total weight of plant, number of pods per plant, 100-seed weight, and grain yield
per plant and harvest index. The parent Bittal-98 was the best general combiner for days
taken to maturity, while parent AUG-786 was the best general combiner for number of
seeds per pod.
The relative magnitude of variation due to GCA and SCA indicated the importance
of additive effects for number of primary branches per plant, plant height, and total weight
of plant. Dominant genetic effects were indicated for days taken to flowering, days taken to
maturity, number of secondary branches per plant, number of pods per plant, number of
seeds per pod, 100-seed weight, grain yield per plant and harvest index. Specific combining
ability effects revealed that it is difficult to select a single good general combiner for major
contributing traits because of negative associations and mutual cancellations, however CM-
98 was good specific combiner for many traits.
108
RAPD analysis showed that two parents Wanhar-2000 and Balksar-2000 were
present in one cluster. The parents Punjab-2000 and Bittal-98 were also present in one
cluster. The results are indicative of the genetic relation. To increase more diversity
breeders need to use parents with diverse origin for future breeding program.
To study inheritance of resistance to wilt, understanding of genetic architecture of
plant and mode of inheritance to disease is important. Six resistant genotypes, one
susceptible and their crosses were studied for the inheritance of wilt. The results indicated
that F1 populations responded to inoculation by formation of necrotic lesion on primary
leaves showing resistance against the disease. Chi-square analysis data segregated into
three classes in F2 and F3 generations showing a close fit of actual to the expected
distribution as 1:2:1 in most of the crosses. The F3 segregation pattern suggested that the
susceptibility was mono-genic in nature and showed dominance of resistance over
susceptibility at single locus. Predominant presence of non-additive gene action for
inheritance of resistance to disease, yield and its parameters suggested that high yielding
and resistant lines may be developed following bi-parental or recurrent selection in early
segregating populations.
Results manifested that biology of yield in chickpea can be understood from yield in
relation to yield. Some balance between yield components results maximum yield. It may
be concluded from present diallelic studies in chickpea genotypes, CM-98 and Bittal-98
showing additive gene effects and better estimates of GCA and SCA would be useful in
development of further promising breeding material. Furthermore, it is suggested that
genetic studies may be supplemented with the modern tools and techniques of genetic
engineering and biotechnology.
109
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